Methods of administering novel integrin activator vaccine compositions

ABSTRACT

Methods for priming of vaccines including administering to a patient a composition including one or more integrin activating compounds capable of adjuvanting an antigen and capable of adjuvanting one or more antigens contained in a vaccine preparation.

RELATED APPLICATIONS

This application is a divisional application of U.S. patent applicationSer. No. 15/140,711 filed Apr. 28, 2016 (28 Apr. 2016), which claimspriority to and the benefit of U.S. Provisional Patent Application Ser.No. 62/154,554 filed Apr. 29, 2015 (29 Apr. 2015).

BACKGROUND OF THE INVENTION 1. Field of the Invention

Embodiments of this invention relates to small molecule integrin ligandmimetics that facilitate integrin-ligand interactions, which may be usedto prepare vaccines, adoptive cell therapies, and immunotherapies forcancer, and a variety of other conditions.

2. Description of the Related Art

There is a major unmet need for a safe and efficacious adjuvant capableof potentiating the immune response in infectious, autoimmune, andallergic diseases, and in other conditions such as cancer. Commonly,adjuvants are compounds or other agents used to enhance the immuneresponse to vaccine antigens (or immunogens). Despite the descriptionsof over one hundred adjuvants in the scientific literature, only alum(aluminum salts or aluminum gels) is currently licensed for use in theUnited States. This is because most adjuvants have unacceptable sideeffect profiles and lack biocompatibility. In fact, even alum, which canbe found in diphtheria-tetanus-pertussis, human papillomavirus, andhepatitis vaccines, can be associated with injection site reactions andis facing increasing scrutiny regarding the potential for cumulativealuminum toxicity.

In the cancer field, there is an unmet need for a safe efficaciousadjuvant capable of boosting the immune response (including bothcellular and humoral) against cancer vaccine antigens. In general,cancer vaccines have been administered without an adjuvant or withspecific cytokines included as adjuvants.

Recombinant, single immunogen cancer vaccines have also been described.One such product which failed in Phase 3 clinical trials is the GVAX®vaccine (Cell Genesys, Inc., South San Francisco, Calif.). This cancervaccine is comprised of a cell line(s) that have been geneticallymodified to secrete granulocyte-macrophage colony stimulating factor(GM-CSF). This cytokine/hormone plays a role in stimulating the body'simmune response to the cancer vaccine. The cells are irradiated forsafety. Cancer vaccination with GVAX® vaccine or other similarapproaches have provided limited benefit over conventional chemotherapy.

Though some studies have utilized specific cytokines as cancer vaccineadjuvants, such as GM-CSF in the GVAX® vaccine, those cytokinestypically enhance only specific features of the immune response, arecostly, and may be unstable outside of very controlled storageconditions. Thus, there is significant need for improvement in the artfor adjuvants displaying increased effectiveness and biocompatibility.

Purified soluble, recombinant and synthetic antigens are often much lessimmunogenic than live or killed whole organism vaccines despite theirbetter tolerability. Thus, the move towards the development of safersubunit vaccines has created a major need for more potent adjuvants. Inparticular, there is an urgent need for adjuvants capable of boostingboth the cellular and/or humoral immune response with more acceptablesafety profiles.

The prerequisites for an ideal cancer adjuvant differ from conventionaladjuvants for many reasons. First, the patients that will receive thevaccines are immuno-compromised because of, for example, impairedmechanisms of antigen presentation, non-responsiveness of activated Tcells and enhanced inhibition of self-reactivity by regulatory T cells.Second, the tumor antigens are usually self-derived in nature, and aretherefore poorly immunogenic. Third, tumors develop escape mechanisms toavoid the immune system, such as tumor immunoediting, which can involvelow or non-expression of MHC class I molecules, and secretion ofsuppressive cytokines. Fourth, even when robust immune responses areelicited to a given tumor antigen, effector functions of the adaptiveresponse can be limited by the activation of immuno-suppressive pathways(CTLA-4/B7, PD-1/PDL-1 axis, IDO1), and effector functions may not belong-lived. Thus, adjuvants for cancer vaccines need to be more potentthan for prophylactic vaccines in not only priming the immune response,but facilitating effector functions, which consequently may be moretoxic and may even induce autoimmune reactions.

As such, there is a clear need for approaches that can selectivelytarget rate limiting steps in the priming of the immune response andpotentiating effector functions. Cellular interactions between integrinsα4β1, α4β7, α5β1, and/or αLβ2 and their cognate ligands are importantmediators of cell-cell adhesion leading to effective priming andeffector functions in the immune system.

SUMMARY OF THE INVENTION

Specifically, the present invention describes compositions and methodsto enhance: (1) the priming of vaccines (including, but not limited to,cancer vaccines); (2) cytolytic activity of adoptive cell therapies(γδT-cells, CTLs, NK, iNKT); (3) immunotherapies (including, but notlimited to, negative checkpoint blockage strategies such as anti-CTLA-4and anti-PD-1 therapies); and (4) biologic therapies (including, but notlimited, to trastuzumab and rituxamab), whereby the mechanism-of-actionincludes antibody dependent cellular cytotoxicity (ADCC).

BRIEF DESCRIPTION OF THE DRAWINGS

The invention can be better understood with reference to the followingdetailed description together with the appended illustrative drawings inwhich like elements are numbered the same:

FIG. 1 depicts C4 enhancement of natural killer cell cytolysis of tumortarget cells. Freshly isolated CD56+ natural killer cells were isolatedfrom the peripheral blood of a healthy volunteer. Cells were eitherincubated with vehicle control (DMSO), or indicated concentrations of C4for 4 h in the presence of tumor target cells (the erythroleukemic cellline K562). Data is presented as a percent increase in tumor cellkilling.

FIG. 2 depicts the natural killer cell line NK-92, which was used atdifferent effector to target ratios in the presence or absence of C4(100 μM). Specific lysis of K562 cells is shown. All data are fromtriplicate determinations, and graphed as the mean±SD.

FIGS. 3A&B depicts activity of C4 in enhancing SDF-la mediated migrationof Jurkat T cell, an exemplary activated T-cell, across membranes coatedwith either VCAM-1 or ICAM-1 (*p<0.05).

FIGS. 4A-C depict a mechanism of ADCC.

DEFINITIONS USED IN THE INVENTION

In addition to having their customary and usual meaning, the followingdefinitions apply where the context permits in the specification andclaims:

“Pharmaceutical composition” refers to a mixture of one or morechemicals, or pharmaceutically acceptable salts thereof, with a suitablecarrier, for administration to a mammal as a medicine.

“Cell therapeutic” refers to a mixture of one or more cells, or one ormore chemicals or pharmaceutically acceptable salts thereof, with asuitable carrier for administration to a mammal as medicine.

“Therapeutically effective amount” refers to that amount of the compoundbeing administered that will relieve at least to some extent one or moreof the symptoms of the disorder being treated. For example, an amount ofthe compound effective to prevent, alleviate or ameliorate symptoms ofdisease or prolong the survival of the subject being treated.

The term “therapeutically ineffective amount” refers to that amount ofthe compound being administered that will not relieve at least to someextent one or more of the symptoms of the disorder being treated. Forexample, doses at which a compound is effective in one individual orpopulation is not effective in other individual or population by meansof intrinsic or acquired drug resistance.

The term “intrinsic resistance” is the innate ability of a cell ororganism to resist activity of a particular a agent through its inherentstructural or functional characteristics, which allow tolerance of aparticular drug or drug class.

The term “acquired resistance” is the adaptive ability of a cell ororganism to resist activity of a particular agent through induction ofinherent structural or functional characteristics, which allow toleranceof a particular drug or drug class.

With respect to a disease or disorder, the term “treatment” refers topreventing, deterring the occurrence of the disease or disorder,arresting, regressing, or providing relief from symptoms or side effectsof the disease or disorder and/or prolonging the survival of the subjectbeing treated.

The term “alkyl” as used herein alone or in combination refers to C₁-C₁₂straight or branched, substituted or unsubstituted saturated chainradicals derived from saturated hydrocarbons by the removal of onehydrogen atom. Representative examples of alkyl groups include methyl,ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, andtert-butyl among others.

The term “alkenyl”, alone or in combination, refers to a substituted orunsubstituted straight-chain or substituted or unsubstitutedbranched-chain alkenyl radical containing from 2 to 10 carbon atoms.Examples of such radicals include, but are not limited to, ethenyl, E-and Z-pentenyl, decenyl and the like.

The term “alkynyl”, alone or in combination, refers to a substituted orunsubstituted straight or substituted or unsubstituted branched chainalkynyl radical containing from 2 to 10 carbon atoms. Examples of suchradicals include, but are not limited to ethynyl, propynyl, propargyl,butynyl, hexynyl, decynyl and the like.

The term “lower” modifying “alkyl”, “alkenyl”, “alkynyl” or “alkoxy”refers to a C₁-C₆ unit for a particular functionality. For example loweralkyl means C₁-C₆ alkyl.

The term “cycloalkyl” as used herein alone or in combination refers to asubstituted or unsubstituted aliphatic ring system having 3 to 10 carbonatoms and 1 to 3 rings, including, but not limited to cyclopropyl,cyclopentyl, cyclohexyl, norbornyl, and adamantyl among others.Cycloalkyl groups can be unsubstituted or substituted with one, two orthree substituents independently selected from lower alkyl, haloalkyl,alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, hydroxy, halo,mercapto, nitro, carboxaldehyde, carboxy, alkoxycarbonyl andcarboxamide. This term is meant to encompass cycloalkenyl andcycloalkynyl groups. “Cycloalkyl” includes cis or trans forms.Furthermore, the substituents may either be in endo or exo positions inthe bridged bicyclic systems.

The term “cycloalkenyl” as used herein alone or in combination refers toa cyclic carbocycle containing from 4 to 8 carbon atoms and one or moredouble bonds. Examples of such cycloalkenyl radicals include, but arenot limited to, cyclopentenyl, cyclohexenyl, cyclopentadienyl and thelike.

The term “cycloalkylalkyl” as used herein refers to a cycloalkyl groupappended to a lower alkyl radical, including, but not limited tocyclohexyl methyl.

The term “halo” or “halogen” as used herein refers to I, Br, Cl or F.

The term “haloalkyl” as used herein refers to a lower alkyl radical, towhich is appended at least one halogen substituent, for examplechloromethyl, fluoroethyl, trifluoromethyl and pentafluoroethyl amongothers.

The term “alkoxy”, alone or in combination, refers to an alkyl etherradical, wherein the term “alkyl” is as defined above. Examples ofsuitable alkyl ether radicals include, but are not limited to, methoxy,ethoxy, n-propoxy, iso-propoxy, n-butoxy, iso-butoxy, sec-butoxy,tert-butoxy and the like.

The term “alkenoxy”, alone or in combination, refers to a radical offormula alkenyl-O—, provided that the radical is not an enol ether,wherein the term “alkenyl” is as defined above. Examples of suitablealkenoxy radicals include, but are not limited to, allyloxy, E- andZ-3-methyl-2-propenoxy and the like.

The term “alkynoxy”, alone or in combination, refers to a radical offormula alkynyl-O—, provided that the radical is not an -ynol ether.Examples of suitable alkynoxy radicals include, but are not limited to,propargyloxy, 2-butynyloxy and the like.

The term “carboxyl” as used herein refers to —CO₂H.

The term “thioalkoxy”, refers to a thioether radical of formulaalkyl-S—, wherein “alkyl” is as defined above.

The term “carboxaldehyde” as used herein refers to —C(O)R, wherein R ishydrogen.

The term “carboxamide” as used herein refers to —C(O)NR₂, wherein R ishydrogen, alkyl or any other suitable substituent.

The term “alkoxyalkoxy” as used herein refers to R_(b) O—R_(c)O—,wherein R_(b) is lower alkyl as defined above and R_(c) is alkylenewherein alkylene is —(CH₂)_(n′)— wherein n′ is an integer from 1 to 6.Representative examples of alkoxyalkoxy groups include methoxymethoxy,ethoxymethoxy, and t-butoxymethoxy among others.

The term “alkylamino” as used herein refers to R_(d) NH—, wherein R_(d)is a lower alkyl group, for example, ethylamino, butylamino, amongothers.

The term “alkenylamino” alone or in combination, refers to a radical offormula alkenyl-NH— or (alkenyl)₂N—, wherein the term “alkenyl” is asdefined above, provided that the radical is not an enamine. An exampleof such alkenylamino radicals is the allylamino radical.

The term “alkynylamino”, alone or in combination, refers to a radical offormula alkynyl-NH— or (alkynyl)₂N—, wherein the term “alkynyl” is asdefined above, provided that the radical is not an amine. An example ofsuch alkynylamino radicals is the propargyl amino radical.

The term “dialkylamino” as used herein refers to R_(e)R_(f)N—, whereinR_(e) and R_(f) are independently selected from lower alkyl, for examplediethylamino, and methyl propylamino, among others.

The term “amino” as used herein refers to H₂N—.

The term “alkoxycarbonyl” as used herein refers to an alkoxyl group aspreviously defined appended to the parent molecular moiety through acarbonyl group. Examples of alkoxycarbonyl include methoxycarbonyl,ethoxycarbonyl, and isopropoxycarbonyl among others.

The term “aryl” or “aromatic” as used herein alone or in combinationrefers to a substituted or unsubstituted carbocyclic aromatic grouphaving about 6 to 12 carbon atoms such as phenyl, naphthyl, indenyl,indanyl, azulenyl, fluorenyl and anthracenyl; or a heterocyclic aromaticgroup selected from the group consisting of furyl, thienyl, pyridyl,pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, 2-pyrazolinyl,pyrazolidinyl, isoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl,1,2,3-triazolyl, 1,3,4-thiadiazolyl, pyridazinyl, pyrimidinyl,pyrazinyl, 1,3,5-triazinyl, 1,3,5-trithianyl, indolizinyl, indolyl,isoindolyl, 3H-indolyl, indolinyl, benzo[b]furanyl,2,3-dihydrobenzofuranyl, benzo[b]thiophenyl, 1H-indazolyl,benzimidazolyl, benzthiazolyl, purinyl, 4H-quinolizinyl, isoquinolinyl,cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 1,8-naphthridinyl,pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl,phenoxyazinyl, pyrazolo[1,5-c]triazinyl and the like. “Arylalkyl” and“alkylaryl” employ the term “alkyl” as defined above. Rings may bemultiply substituted. Aromatic rings may be fused with other aromatic ornon-aromatic rings to form multicyclic rings, and are also encompassedby the term “aromatic,” as used herein.

The term “aralkyl”, alone or in combination, refers to an arylsubstituted alkyl radical, wherein the terms “alkyl” and “aryl” are asdefined above. Examples of suitable aralkyl radicals include, but arenot limited to, phenylmethyl, phenethyl, phenylhexyl, diphenylmethyl,pyridylmethyl, tetrazolyl methyl, furylmethyl, imidazolyl methyl,indolylmethyl, thienylpropyl and the like.

The term “aralkenyl”, alone or in combination, refers to an arylsubstituted alkenyl radical, wherein the terms “aryl” and “alkenyl” areas defined above.

The term “arylamino”, alone or in combination, refers to a radical offormula aryl-NRg-, wherein “aryl” is as defined above. Rg may beselected from the group consisting of H, lower alkyl, aryl and aralkylamong others. Examples of arylamino radicals include, but are notlimited to, phenylamino(anilido), naphthlamino, 2-, 3-, and4-pyridylamino and the like.

The term “biaryl”, alone or in combination, refers to a radical offormula aryl-aryl, wherein the term “aryl” is as defined above.

The term “thioaryl”, alone or in combination, refers to a radical offormula aryl-S—, wherein the term “aryl” is as defined above. An exampleof a thioaryl radical is the thiophenyl radical.

The term “aroyl”, alone or in combination, refers to a radical offormula aryl-CO—, wherein the term “aryl” is as defined above. Examplesof suitable aromatic acyl radicals include, but are not limited to,benzoyl, 4-halobenzoyl, 4-carboxybenzoyl, naphthoyl, pyridylcarbonyl andthe like.

The term “heterocyclyl”, alone or in combination, refers to anon-aromatic 3- to 10-membered ring containing at least one endocyclicN, O, or S atom. The heterocycle may be optionally aryl-fused. Theheterocycle may also optionally be substituted with at least onesubstituent which is independently selected from the group consisting ofhydrogen, halogen, hydroxyl, amino, nitro, trifluoromethyl,trifluoromethoxy, alkyl, aralkyl, alkenyl, alkynyl, aryl, cyano,carboxyl, alkoxycarbonyl, carboxyalkyl, oxo, arylsulfonyl andaralkylaminocarbonyl among others.

The term “alkylheterocyclyl” as used herein refers to an alkyl group aspreviously defined appended to the parent molecular moiety through aheterocyclyl group.

The term “heterocyclylalkyl” as used herein refers to a heterocyclylgroup as previously defined appended to the parent molecular moietythrough an alkyl group.

The term “aminal” as used herein refers to a hemi-acetal of thestructure RCH(NH₂)(OH).

The terms “electron-withdrawing” or “electron-donating” refer to theability of a substituent to withdraw or donate electrons relative tothat of hydrogen if hydrogen occupied the same position in the molecule.These terms are well-understood by one skilled in the art and arediscussed in ADVANCED ORGANIC CHEMISTRY by J. March, 1985, pp. 16-18,incorporated herein by reference. Electron withdrawing groups includehalo, nitro, carboxyl, lower alkenyl, lower alkynyl, carboxaldehyde,carboxyamido, aryl, quaternary ammonium, trifluoromethyl, and aryl loweralkanoyl among others. Electron donating groups include such groups ashydroxy, lower alkyl, amino, lower alkylamino, di(lower alkyl)amino,aryloxy, mercapto, lower alkylthio, lower alkylmercapto, and disulfideamong others. One skilled in the art will appreciate that the aforesaidsubstituents may have electron donating or electron withdrawingproperties under different chemical conditions. Moreover, the presentinvention contemplates any combination of substituents selected from theabove-identified groups.

The most preferred electron donating or electron withdrawingsubstituents are halo, nitro, alkanoyl, carboxaldehyde, arylalkanoyl,aryloxy, carboxyl, carboxamide, cyano, sulfonyl, sulfoxide,heterocyclyl, guanidine, quaternary ammonium, lower alkenyl, loweralkynyl, sulfonium salts, hydroxy, lower alkoxy, lower alkyl, amino,lower alkylamino, di(lower alkyl)amino, amine lower alkyl mercapto,mercaptoalkyl, alkylthio and alkyldithio.

Use of the above terms is meant to encompass substituted andunsubstituted moieties. Substitution may be by one or more groups suchas alcohols, ethers, esters, amides, sulfones, sulfides, hydroxyl,nitro, cyano, carboxy, amines, heteroatoms, lower alkyl, lower alkoxy,lower alkoxycarbonyl, alkoxyalkoxy, acyloxy, halogens, trifluoromethoxy,trifluoromethyl, alkyl, aralkyl, alkenyl, alkynyl, aryl, cyano, carboxy,carboalkoxy, carboxyalkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl,alkylheterocyclyl, heterocyclylalkyl, oxo, arylsulfonyl andaralkylaminocarbonyl or any of the substituents of the precedingparagraphs or any of those substituents either attached directly or bysuitable linkers. The linkers are typically short chains of 1-3 atomscontaining any combination of —C—, —C(O)—, —NH—, —S—, —S(O)—, —O—,—C(O)O— or —S(O)O—. Rings may be substituted multiple times.

The term “mammals” includes humans and other animals.

The term “heteroatom” as used herein encompasses nitrogen, sulfur andoxygen.

The term “alpha” as used herein indicates the position immediatelyadjacent to the position described.

The term “inactive ingredient” as used herein indicated a harmless drugthat is ordinarily used as an inactive ingredient, such as a coloring,emulsifier, excipient, flavoring, lubricant, preservative, or solvent,in the preparation of other drugs shall be exempt from section 502(f)(1)of the act (21 CFR 201.117).

The term “excipient” as used herein means any substance other than theactive drug or product which has been appropriately evaluated for safetyand is included in a drug delivery system to either aid the processingof the drug delivery system during its manufacture; protect, support, orenhance stability, bioavailability, or patient acceptability; assist inproduct identification; or enhance any other attribute of the overallsafety and effectiveness of the drug delivery system during storage oruse (40 CFR 63.1251).

The term “effector cell” as used herein means a cell that can bind toand either engulf or induce cytolysis of a target cell. Effector celltypes may include, but are not limited to: 1) a cytotoxic T-lymphocytethat binds to target cells and is activated by an antigen specific Tcell receptor, 2) monocytes, macrophage, natural killer (NK) cells, andneutrophils that bind to and lyse target cells through interactions offragment crystallizable (Fc) receptors and monoclonal antibody opsonizedtarget cells, 3) NK and NK-variants that bind to and lyse target cellsindependent of antigen specificity, 4) Tumor Infiltrating Lymphocytes(TILs) which are lymphocytes isolated from tumors and expanded ex vivothat express cell surface markers including, but not limited to CD3,CD8, or CD4, 5) T cells genetically engineered with tumor specific Tcell receptors or chimeric antigen receptors that possess cell surfacemarkers including but not limited to CD3, CD8, or CD4.

The term “adoptive T cell” is a effector cell that is derived from anaive T cell or activated T cell capable of effector functions.

The term “solid tumor” as used herein means an abnormal mass of tissuethat usually does not contain cysts or liquid areas. Solid tumors may bebenign (not cancer), or malignant (cancer). Different types of solidtumors are named for the type of cells that form them. Examples of solidtumors are sarcomas, carcinomas, and lymphomas.

The term “small molecule agonist” as used herein is not a conventionalligand, and is synonymous to a stabilizer of a cognate ligand-receptorinteraction.

Abbreviations Used in the Invention

The following abbreviations are used herein: Ac is acetyl, AcOH isacetic acid, ADCC is antibody dependent cellular cytotoxicity, 6-Ahx-OHis 6-aminohexanoic acid, APC is antigen presenting cell, BATDA is bis(acetoxymethyl) 2,2′:6′,2″-terpyridine-6,6″-dicarboxylate, BCG isbacillus calmette-guérin, Bn is benzyl, Boc is tert-butyloxycarbonyl,nBu is n-butyl, nBuLi is n-butyllithium, 1.6M in hexanes (unless otherconcentration noted), Cbz is benzyloxycarbonyl, CD is cluster ofdifferentiation, CDI is N,N′-carbonyldiimidazole, COMU is(1-cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbeniumhexafluorophosphate, CTL is cytotoxic T lymphocyte, CTLA-4 is cytotoxicT lymphocyte-associated antigen-4, Dab is 2,4-diaminobutyryl, DBU is1,8-diazabicyclo[5.4.0]undec-7-ene, DCE is 1,2-dichloroethane, DCHA isdicyclohexylamine, DCM is dichloromethane (methlyene chloride), dioxaneis 1,4-dioxane, DIPEA is N,N-diisopropylethylamine, DMED isN,N′-dimethylethylene diamine, DMF is N,N-dimethylformamide, DMSO isdimethylsulfoxide Et is ethyl, EtOH is ethanol, FBS is fetal bovineserum, Fc is fragment crystallizable, FGF is fibroblast growth factor,Fmoc is 9H-fluoren-9-ylmethyloxycarbonyl, G-CSF is granulocyte colonystimulating factor, Glu is glutamic acid, Gly is glycine, HBTU isO-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate,HMDS is hexamethyldisilazane, ICAM-1 is intercellular adhesionmolecule-1, IDO is indoleamine 2,3-dioxygenase, iNKT is invariantnatural killer T cell, iPr is isopropyl, KHMDS is potassiumbis(trimethylsilyl)amide, LFA is lymphocyte function-associated antigenwhere LFA-1 is the integrin αLβ2, Lys is lysine, LHMDS is lithiumbis(trimethylsilyl)amide, MAdCAM-1 is mucosal addressin cell adhesionmolecule-1, Me is methyl, MeOH is methanol, MHC is majorhistocompatibility complex, NK is natural killer, Nle is norleucine, NMMis 4-methylmorpholine, NSMC is N-succinimidyl-N-methylcarbamate, OAc isacetate, Orn is Ornithine, PBS is phosphate buffered saline, PD1 isprogrammed death 1, PDL1 is programmed death ligand 1, pTsOH ispara-toluenesulfonic acid, Ph is phenyl, RT is room temperature, SDF1αis stromal cell derived factor-1α, tBu is tert-butyl, TBS istris-buffered saline, TDA is 2,2′:6′,2″-terpyridine-6,6″-dicarboxylicacid, TEA is triethylamine, TIL is tumor infiltrating lymphocyte, Tfa istrifluoroacetyl, Th1 is T helper 1, Th2 is T helper 2, THF istetrahydrofuran, Tol is toluene, Tyr is tyrosine, VCAM-1 is vascularcell adhesion molecule-1, Veh is vehicle, VLA is very late activationantigen where VLA-4 is the integrin α4β1 and VLA-5 is the integrin α5β1,and Z is benzyloxycarbonyl.

DETAILED DESCRIPTION OF THE INVENTION

We have invented small molecule integrin ligand mimetics that facilitateintegrin-ligand interactions which can be used to prepare vaccines,adoptive cell therapies, and immunotherapies for cancer, and a varietyof other conditions. The inventors have found that certain smallmolecules or chemical compounds are capable of enhancing the therapeuticefficacy of vaccines, adoptive cell therapies, immunotherapies forcancer, antibody dependent cellular cytotoxicity (ADCC), and/or avariety of other conditions.

In certain embodiments, the present invention relates to pharmaceuticalpreparations of a vaccine comprising an antigen, and an effective amountof one or a plurality of enhancing compounds, where the enhancingcompounds are capable of enhancing integrin-mediated binding ofintegrins of a cell to their respective ligand, where the integrinsinclude 4β1, α4β7, 5β1, and/or αLβ2, where the ligands include VCAM-1,fibronectin, MAdCAM-1, ICAM-1, and/or ICAM-2, where the antigen is apurified protein, a purified peptide, a cell, or a cell lysate. In otherembodiments, the preparation is an anti-cancer vaccine. In otherembodiments, the pharmaceutical preparation of claim 1, furthercomprising an adjuvant comprising a non-specific adjuvant substanceand/or a specific adjuvant substance, where the substances are capableof eliciting an immune response in response to the antigen. In otherembodiments, the non-specific adjuvant substance is selected from thegroup consisting of BCG, complete freund's adjuvant, alum, and/ornoscapine, and the specific adjuvant substance is selected from thegroup consisting of G-CSF, FGF, Toll-like receptor agonists, and/orimmune checkpoint inhibitors targeting proteins selected from the groupconsisting of CTL-4, PD-1, PDL-1, and/or IDO-1. In certain embodiments,the enhancing compounds are specific adjuvants.

In certain embodiments, the present invention relates methods to treat apatient with an effective amount of one or a plurality of enhancingcompounds, wherein the associated enhancing compounds are capable ofenhancing integrin-mediated binding of integrins of a cell to theirrespective ligand, wherein the integrins include α4β1, α4β7, α5β1,and/or αLβ2, wherein the ligands include VCAM-1, fibronectin, MAdCAM-1,ICAM-1, and/or ICAM-2, in combination with a therapeutically ineffectivedose of immune checkpoint inhibitor, cytotoxic chemotherapy, cancervaccine, wherein the patient has been diagnosed with cancer.

In certain embodiments, the present invention relates methods to treat apatient with an effective amount of one or a plurality of enhancingcompounds, wherein the associated enhancing compounds are capable ofenhancing integrin-mediated binding of integrins of a cell to theirrespective ligand, wherein the integrins include α4β1, α4β7, α5β1,and/or αLβ2, wherein the ligands include VCAM-1, fibronectin, MAdCAM-1,ICAM-1, and/or ICAM-2, in combination with a therapeutically ineffectivedose of immune checkpoint inhibitor, cytotoxic chemotherapy, cancervaccine, wherein the patient has been diagnosed with solid tumors in thelung, prostate, breast, colon, skin, brain, or pancreas, wherein theenhancing compound is intratumorally injected, locally infused tospecific organs or tissues systemically administered by enteral orparenteral route of administration.

In certain embodiments, the present invention relates methods to treat apatient with an effective amount of one or a plurality of enhancingcompounds, where the associated enhancing compounds are capable ofenhancing interin-mediated binding of integrins of a cell to theirrespective ligand, wherein the integrins include α4β1, α4β7, α5β1,and/or αLβ2, wherein the ligands include VCAM-1, fibronectin, MAdCAM-1,ICAM-1, ICAM-2, and/or vitronectin, in combination with atherapeutically ineffective dose of immune checkpoint inhibitor,cytotoxic chemotherapy, wherein the patient has been diagnosed withhematologic cancers, wherein the integrin mimetic is systemicallyadministered by enteral or parentral route of administration.

In certain embodiments, the present invention relates pharmaceuticalpreparations comprising NK cells, activated NK cells, engineered NKcells, or NK cell lines (and derivatives thereof) treated with aneffective amount of one or a number of compounds capable of activatingintegrin mediated interactions with their cognate ligands.

In certain embodiments, the present invention relates methods to treat apatient with a pharmaceutical preparation comprising NK cells, activatedNK cells, engineered NK cells, or NK cell lines (and derivativesthereof) delivered in combination with (either prior to, during, orafter) an effective amount of one or a number of compounds capable ofactivating integrin mediated interactions with their cognate ligands.

In certain embodiments, the present invention relates methods to treat apatient with a pharmaceutical preparation comprising a therapeuticallyineffective amount of NK cells, activated NK cells, engineered NK cells,or NK cell lines (and derivatives thereof) delivered in combination with(either prior to, during, or after) an effective amount of one or anumber of compounds capable of activating integrin mediated interactionswith their cognate ligands.

In certain embodiments, the present invention relates pharmaceuticalpreparations comprising invariant natural killer T (iNKT) cells,activated iNKT cells, engineered iNKT cells, or iNKT cell lines (andderivatives thereof) treated with an effective amount of one or a numberof compounds capable of activating integrin mediated interactions withtheir cognate ligands.

In certain embodiments, the present invention relates methods to treat apatient with a pharmaceutical preparation comprising iNKT cells,activated iNKT cells, engineered iNKT cells, or iNKT cell lines (andderivatives thereof) delivered in combination with (either prior to,during, or after) an effective amount one or a number of compoundscapable of activating integrin mediated interactions with their cognateligands.

In certain embodiments, the present invention relates methods to treat apatient with a pharmaceutical preparation comprising a therapeuticallyineffective amount of iNKT cells, activated iNKT cells, engineered iNKTcells, or iNKT cell lines (and derivatives thereof) delivered incombination with (either prior to, during, or after) an effective amountone or a number of compounds capable of activating integrin mediatedinteractions with their cognate ligands.

In certain embodiments, the present invention relates methodspharmaceutical preparations comprising γδT cells, activated iNKT cells,engineered γδT cells, or γδT cell lines (and derivatives thereof)treated with an effective amount of one or a number of compounds capableof activating integrin mediated interactions with their cognate ligands.

In certain embodiments, the present invention relates methods to treat apatient with a pharmaceutical preparation comprising γδT cells,activated iNKT cells, engineered γδT cells, or γδT cell lines (andderivatives thereof) delivered in combination with (either prior to,during, or after) an effective amount one or a number of compoundscapable of activating integrin mediated interactions with their cognateligands.

In certain embodiments, the present invention relates methods to treat apatient with a pharmaceutical preparation comprising γδT cells,activated iNKT cells, engineered γδT cells, or γδT cell lines (andderivatives thereof) delivered in combination with (either prior to,during, or after) an effective amount one or a number of compoundscapable of activating integrin mediated interactions with their cognateligands.

In certain embodiments, the present invention relates pharmaceuticalpreparations comprising cytotoxic T lymphocytes (CTLs) (including, butnot limited to, tumor infiltrating lymphocytes, lymphocytes expanded inan antigen specific manner, lymphocytes engineered to express chimericantigen receptors) treated with an effective amount of one or a numberof compounds capable of activating integrin mediated interactions withtheir cognate ligands for the purpose of enhancing CTL tumoricidalactivity.

In certain embodiments, the present invention relates methods to treat apatient with a pharmaceutical preparation comprising cytotoxic Tlymphocytes (CTLs) (including, but not limited to, tumor infiltratinglymphocytes, lymphocytes expanded in an antigen specific manner,lymphocytes engineered to express chimeric antigen receptors) deliveredin combination with (either prior to, during, or after) an effectiveamount of one or a number of compounds capable of activating integrinmediated interactions with their cognate ligands for the purpose ofenhancing CTL tumoricidal activity.

In certain embodiments, the present invention relates methods to treat apatient with a pharmaceutical preparation comprising a therapeuticallyineffective amount of cytotoxic T lymphocytes (CTLs) (including, but notlimited to, tumor infiltrating lymphocytes, lymphocytes expanded in anantigen specific manner, lymphocytes engineered to express chimericantigen receptors) delivered in combination with (either prior to,during, or after) an effective amount of one or a number of compoundscapable of activating integrin mediated interactions with their cognateligands for the purpose of enhancing CTL tumoricidal activity.

In certain embodiments, the present invention relates methods to treat apatient with a therapeutic antibody, wherein the antibody contains a Fcregion, wherein the antibody is, without limitation, trastuzumab,cetuximab, ipilimumab, nivolumab rituximab, alemtuzumab, atumumab, ortositumomab, and an effective amount of one or a number of compoundscapable of activating integrin mediated interactions with their cognateligands.

In certain embodiments, the present invention relates methods to treat apatient with therapeutically ineffective amount of therapeutic antibody,wherein the antibody contains a Fc region, wherein the antibody is,without limitation, trastuzumab, cetuximab, ipilimumab, nivolumabrituximab, alemtuzumab, atumumab, or tositumomab, and an effectiveamount of one or a number of compounds capable of activating integrinmediated interactions with their cognate ligands.

In certain embodiments, the present invention relates pharmaceuticalcompositions comprising a therapeutically ineffective amount oftherapeutic antibody, wherein the antibody contains a Fc fragment and aneffective amount of one or a number of compounds capable of activatingintegrin mediated interactions with their cognate ligands.

In certain embodiments, the chemical compounds are given by the generalFormula (I):

R¹-M¹-N(R²)-M²-M³-M⁴-M⁵-M⁶-R³  (I)

where:

-   -   R¹ is selected from the group consisting of aryl and aralkyl,    -   R² is alkyl, aryl, or aralkyl,    -   M¹ is CH₂,    -   M² is CO,    -   M³ is O, S, or NR⁶, where R⁶ when present is hydrogen or lower        alkyl,    -   M⁴ is absent or CH₂,    -   M⁵ is (CR¹¹R¹²), where R¹¹ is hydrogen, R¹² is selected from the        group consisting of hydrogen, NR²¹CONR²²R²³, NR²¹COR²⁴,        NR²¹SO₂R²⁴, NR²¹COOR²⁴, OCOR²⁴, OR²⁴, O(CH₂CH₂O)_(s)R²⁴, COOR²⁴,        alkyl, and hydroxyalkyl, where s is an integer of 1 to 6, R²¹        and R²² when present are independently selected from the group        consisting of hydrogen or lower alkyl, R²³ when present is        selected from the group consisting of hydroxyalkyl, alkoxyalkyl,        alkyl, aryl, aralkyl and alkoxycarbonylalkyl, provided that when        M³ is NR⁶, M⁴ is absent, and R¹² is CONR²²R²³, then R²³ is not        1-(1,3-benzodioxol-5-yl)-3-ethoxy-3-oxopropyl, R²⁴ when present        is selected from the group consisting of alkyl, aryl, aralkyl,        heterocyclyl, cycloalkyl, cycloalkylalkyl, and        heterocyclylalkyl,    -   M⁶ is (CH₂)_(q), wherein q is an integer from 0 to 6,    -   R³ is selected from the group consisting of hydrogen, CONR¹³R¹⁴,        NR¹⁵COOR¹⁶, NR¹⁵COR¹⁶, NR¹⁵CONR¹³R¹⁴, NR¹⁵SO₂R¹⁶, OCOR¹⁶,        COOR¹⁶, OR¹⁶, SR¹⁶, heterocyclyl, hydroxyl, hydroxyalkyl,        guanadino, alkyl and aryl, where R¹³ and R¹⁵ when present are        independently hydrogen or lower alkyl, R¹⁴ and R¹⁶ when present        are independently selected from the group consisting of        hydrogen, alkyl, aryl, aralkyl, cycloalkyl, heterocyclyl,        cycloalkylalkyl, and heterocyclylalkyl,    -   R¹, R², R³, R¹², R¹⁴, R¹⁶, R²³ and R²⁴ when present may        independently be either unsubstituted or substituted with one or        more substituents selected from the group consisting of alkyl,        aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl,        heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy, azido,        haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl,        halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino,        arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl),        —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl),        —NHSO₂(aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl,        —OCO(alkylamino), and —OCO(dialkylamino).

Embodiments of the present invention provide methods to increase thepotency and or efficacy of effector cells by increasing theirintratumoral bioavailability, where the methods include the steps ofsuffusing a composition directly into a target tissue, or systemicallyinfusing into of a patient, where the composition includes effectorcells treated with an effective amount of one or a plurality of chemicalcompounds capable of enhancing integrin-mediated binding of cells totheir respective ligands and an additional amount of one or a pluralityof chemical compounds capable of enhancing integrin-mediated binding ofcells to their respective ligands. In certain embodiments, integrinstargeted by these compounds include, but are not limited to, α4β1, α4β7,α5β1, αLβ2 and αVβ3. In various embodiments, ligands include, but arenot limited to, VCAM-1, fibronectin, MAdCAM-1, ICAM-1, ICAM-2, andvitronectin.

In other embodiments, the chemical compounds are given by the generalFormula (I):

R¹-M¹-N(R²)-M²-M³-M⁴-M⁵-M⁶-R³  (I)

where:

-   -   R¹ is selected from the group consisting of aryl and aralkyl,    -   R² is alkyl, aryl, or aralkyl,    -   M¹ is CH₂,    -   M² is CO,    -   M³ is O, S, or NR⁶, where R⁶ when present is hydrogen or lower        alkyl,    -   M⁴ is absent or CH₂,    -   M⁵ is (CR¹¹R¹²), where R¹¹ is hydrogen, R¹² is selected from the        group consisting of hydrogen, NR²¹CONR²²R²³, NR²¹COR²⁴,        NR²¹SO₂R²⁴, NR²¹COOR²⁴, OCOR²⁴, OR²⁴, O(CH₂CH₂O)_(s)R²⁴, COOR²⁴,        alkyl, and hydroxyalkyl, where s is an integer of 1 to 6, R²¹        and R²² when present are independently selected from the group        consisting of hydrogen or lower alkyl, R²³ when present is        selected from the group consisting of hydroxyalkyl, alkoxyalkyl,        alkyl, aryl, aralkyl and alkoxycarbonylalkyl, provided that when        M³ is NR⁶, M⁴ is absent, and R¹² is CONR²²R²³, then R²³ is not        1-(1,3-benzodioxol-5-yl)-3-ethoxy-3-oxopropyl, R²⁴ when present        is selected from the group consisting of alkyl, aryl, aralkyl,        heterocyclyl, cycloalkyl, cycloalkylalkyl, and        heterocyclylalkyl,    -   M⁶ is (CH₂)_(q), wherein q is an integer from 0 to 6,    -   R³ is selected from the group consisting of hydrogen, CONR¹³R¹⁴,        NR¹⁵COOR¹⁶, NR¹⁵COR¹⁶, NR¹⁵CONR¹³R¹⁴, NR¹⁵SO₂R¹⁶, OCOR¹⁶,        COOR¹⁶, OR¹⁶, SR¹⁶, heterocyclyl, hydroxyl, hydroxyalkyl,        guanadino, alkyl and aryl, where R¹³ and R¹⁵ when present are        independently hydrogen or lower alkyl, R¹⁴ and R¹⁶ when present        are independently selected from the group consisting of        hydrogen, alkyl, aryl, aralkyl, cycloalkyl, heterocyclyl,        cycloalkylalkyl, and heterocyclylalkyl,    -   R¹, R², R³, R¹², R¹⁴, R¹⁶, R²³ and R²⁴ when present may        independently be either unsubstituted or substituted with one or        more substituents selected from the group consisting of alkyl,        aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl,        heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy, azido,        haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl,        halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino,        arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl),        —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl),        —NHSO₂(aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl,        —OCO(alkylamino), and —OCO(dialkylamino).

In other embodiments, the chemical compounds are given by the generalformula (I):

R¹-M¹-N(R²)-M²-M³-M⁴-M⁵-M⁶-R³  (I)

where R¹ is selected from the group consisting of aryl and aralkyl, R²is alkyl, aryl, or aralkyl, M¹ is CH₂, M² is CO, M³ is O, S, or NR⁶,where R⁶ when present is hydrogen or lower alkyl, M⁴ is absent or CH₂,M⁵ is (CR¹¹R¹²), where R¹¹ is hydrogen, R¹² is selected from the groupconsisting of hydrogen, NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂OR²⁴,NR²¹SO₂R²⁴, NR²¹COOR²⁴, OCOR²⁴, OR²⁴, O(CH₂CH₂O)_(s)R²⁴, COOR²⁴, alkyl,and hydroxyalkyl, where s is an integer of 1 to 6, R²¹ and R²² whenpresent are independently selected from the group consisting of hydrogenor lower alkyl, R²³ when present is selected from the group consistingof hydroxyalkyl, alkoxyalkyl, alkyl, aryl, aralkyl andalkoxycarbonylalkyl, provided that when M³ is NR⁶, M⁴ is absent, thenR²³ is not 1-(1,3-benzodioxol-5-yl)-3-ethoxy-3-oxopropyl, R²⁴ whenpresent is selected from the group consisting of alkyl, aryl, aralkyl,heterocyclyl, cycloalkyl, cycloalkylalkyl, and heterocyclylalkyl, andmixtures or combinations thereof, M⁶ is (CH₂)_(q), wherein q is aninteger from 0 to 6, R³ is selected from the group consisting ofhydrogen, CONR¹³R¹⁴, NR¹⁵COOR¹⁶, NR¹⁵COR¹⁶, NR¹⁵CONR¹³R¹⁴, NR¹⁵SO₂R¹⁶,OCOR¹⁶, COOR¹⁶, OR¹⁶, SR¹⁶, heterocyclyl, hydroxyl, hydroxyalkyl,guanadino, alkyl and aryl, where R¹³ and R¹⁵ when present areindependently hydrogen or lower alkyl, R¹⁴ and R¹⁶ when present areindependently selected from the group consisting of hydrogen, alkyl,aryl, aralkyl, cycloalkyl, heterocyclyl, cycloalkylalkyl, andheterocyclylalkyl, R¹, R², R³, R¹², R¹⁴, R¹⁶, R²³ and R²⁴ when presentmay independently be either unsubstituted or substituted with one ormore substituents selected from the group consisting of alkyl, aryl,aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl,heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl,aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino,alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl),—NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO₂(alkyl),—NHSO₂(aryl), —NHSO₂(aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl,—OCO(alkylamino), —OCO(dialkylamino), and mixtures or combinationsthereof. In other embodiments, the compounds have R¹ is aryl or aralkyl,R² is alkyl or aralkyl, M¹ is CH₂, M² is CO, M³ is absent or is O orCH₂, M⁴ is absent or is CH₂, M⁵ is absent or is O or (CR¹¹R¹²), R¹¹ ishydrogen, R¹² is selected from the group consisting of hydrogen,NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴ and NR²¹COOR²⁴, M⁶ is selected fromthe group consisting of (CH₂)_(q), (CH₂)_(q)—CH═CH—(CH₂)_(r),(CH₂)_(q)-arylene-(CH₂)_(r) and (CH₂CH₂O)_(q), q and r are independentlyintegers from 0 to 6, R³ is CONR¹³R¹⁴, R²¹ and R²² each of which, whenpresent is independently selected from the group of hydrogen and loweralkyl, R¹³, R¹⁴, R²³ and R²⁴, each of which, when present isindependently selected from the group consisting of hydrogen, alkyl,aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl andaralkyl, and R¹, R², R¹³, R¹⁴, R²³ and R²⁴ when present may be eitherunsubstituted or substituted with one or more substituents selected fromthe group consisting of alkyl, aryl, aralkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl,hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl,alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino,arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl),—NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl), —NHSO₂(aralkyl),alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino),—OCO(dialkylamino), and mixtures or combinations thereof. In otherembodiments, the compounds have R¹ is aryl or aralkyl, R² is alkyl oraralkyl, M¹ is CH₂, M² is SO₂ or CO, M³ is absent or is CH₂, M⁴ isabsent or is CH₂, M⁵ is absent or is (CR¹¹R¹²), R¹¹, when present, ishydrogen, R¹², when present, is selected from the group consisting ofhydrogen, alkyl, NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴ and NR²¹COOR²⁴, M⁶is (CH₂)_(q), or NR³⁴(CH₂)_(q), q is an integer from 0 to 6, R³ isselected from the group consisting of CONR¹³R¹⁴, SO₂NR¹³R¹⁴, NR¹⁵COOR¹⁶,NR¹⁵COR¹⁶, NR¹⁵CONR¹³R¹⁴, and NR¹⁵SO₂R¹⁶, R¹⁵, R¹⁶, R²¹ and R²², each ofwhich when present, is independently selected from the group ofhydrogen, lower alkyl, and aralkyl, R¹³, R¹⁴, R²³ and R²⁴, each ofwhich, when present is independently selected from the group consistingof hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl,heterocyclylalkyl and aralkyl, R³⁴, when present, is selected form thegroup consisting of alkyl, aralkyl, COR³⁵, and SO₂R³⁵, R³⁵ when present,is selected form the group consisting of alkyl, aryl, and aralkyl, andR¹, R², R¹³, R¹⁴, R¹⁵, R¹⁶, R²³, R²⁴, R³⁴ and R³⁵, when present, may beeither unsubstituted or substituted with one or more substituentsselected from the group consisting of alkyl, aryl, aralkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl,hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl,alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino,arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl),—NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl), —NHSO₂(aralkyl),alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino), and—OCO(dialkylamino), with the proviso that when M² is CO, then M⁶ isNR³⁴(CH₂)_(q), wherein q is not 0. In other embodiments, the chemicalcompounds are inactive ingredients or excipients. In other embodiments,the effective amount in carrier is greater than 1 fM and less than 100nM. In other embodiments, the effective amount in carrier is greaterthan 1 fM and less than 50 nM. In other embodiments, the effectiveamount in carrier is greater than 1 fM and less than 25 nM.

In other embodiments, the chemical compounds are given by the generalformula (I):

R¹-M¹-N(R²)-M²-M³-M⁴-M⁵-M⁶-R³  (I)

where R¹ is selected from the group consisting of aryl and aralkyl, R²is alkyl, aryl, or aralkyl, M¹ is CH₂, M² is CO, M³ is O, S, or NR⁶,where R⁶ when present is hydrogen or lower alkyl, ⁴ is absent or CH₂, M⁵is (CR¹¹R¹²), where R¹¹ is hydrogen, R¹² is selected from the groupconsisting of hydrogen, NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴,NR²¹COOR²⁴, OCOR²⁴, OR²⁴, O(CH₂CH₂O)_(s)R²⁴, COOR²⁴, alkyl, andhydroxyalkyl, where s is an integer of 1 to 6, R²¹ and R²² when presentare independently selected from the group consisting of hydrogen orlower alkyl, R²³ when present is selected from the group consisting ofhydroxyalkyl, alkoxyalkyl, alkyl, aryl, aralkyl and alkoxycarbonylalkyl,provided that when M³ is NR⁶, M⁴ is absent, then R²³ is not1-(1,3-benzodioxol-5-yl)-3-ethoxy-3-oxopropyl, R²⁴ when present isselected from the group consisting of alkyl, aryl, aralkyl,heterocyclyl, cycloalkyl, cycloalkylalkyl, and heterocyclylalkyl, andmixtures or combinations thereof, M⁶ is (CH₂)_(q), wherein q is aninteger from 0 to 6, R³ is selected from the group consisting ofhydrogen, CONR¹³R¹⁴, NR¹⁵COOR¹⁶, NR¹⁵COR¹⁶, NR¹⁵CONR¹³R¹⁴, NR¹⁵SO₂R¹⁶,OCOR¹⁶, COOR¹⁶, OR¹⁶, SR¹⁶, heterocyclyl, hydroxyl, hydroxyalkyl,guanadino, alkyl and aryl, where R¹³ and R¹⁵ when present areindependently hydrogen or lower alkyl, R¹⁴ and R¹⁶ when present areindependently selected from the group consisting of hydrogen, alkyl,aryl, aralkyl, cycloalkyl, heterocyclyl, cycloalkylalkyl, andheterocyclylalkyl, R¹, R², R³, R¹², R¹⁴, R¹⁶, R²³ and R²⁴ when presentmay independently be either unsubstituted or substituted with one ormore substituents selected from the group consisting of alkyl, aryl,aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl,heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl,aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino,alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl),—NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO₂(alkyl),—NHSO₂(aryl), —NHSO₂(aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl,—OCO(alkylamino), —OCO(dialkylamino), and mixtures or combinationsthereof. In other embodiments, the chemical compounds have R¹ is aryl oraralkyl, R² is alkyl or aralkyl, M¹ is CH₂, M² is CO, M³ is absent or isO or CH₂, M⁴ is absent or is CH₂, M⁵ is absent or is O or (CR¹¹R¹²), R¹¹is hydrogen, R¹² is selected from the group consisting of hydrogen,NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴ and NR²¹COOR²⁴, M⁶ is selected fromthe group consisting of (CH₂)_(q), (CH₂)_(q)—CH═CH—(CH₂)_(r),(CH₂)_(q)-arylene-(CH₂)_(r) and (CH₂CH₂O)_(q), q and r are independentlyintegers from 0 to 6, R³ is CONR¹³R¹⁴, R²¹ and R²² each of which, whenpresent is independently selected from the group of hydrogen and loweralkyl, R¹³, R¹⁴, R²³ and R²⁴, each of which, when present isindependently selected from the group consisting of hydrogen, alkyl,aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl andaralkyl, and R¹, R², R¹³, R¹⁴, R²³ and R²⁴ when present may be eitherunsubstituted or substituted with one or more substituents selected fromthe group consisting of alkyl, aryl, aralkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl,hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl,alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino,arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl),—NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl), —NHSO₂(aralkyl),alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino),—OCO(dialkylamino), and mixtures or combinations thereof. In otherembodiments, the chemical compounds have R¹ is aryl or aralkyl, R² isalkyl or aralkyl, M¹ is CH₂, M² is SO₂ or CO, M³ is absent or is CH₂, M⁴is absent or is CH₂, M⁵ is absent or is (CR¹¹R¹²), R¹¹, when present, ishydrogen, R¹², when present, is selected from the group consisting ofhydrogen, alkyl, NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴ and NR²¹COOR²⁴, M⁶is (CH₂)_(q), or NR³⁴(CH₂)_(q), q is an integer from 0 to 6, R³ isselected from the group consisting of CONR¹³R¹⁴, SO₂NR¹³R¹⁴, NR¹⁵COOR¹⁶,NR¹⁵COR¹⁶, NR¹⁵CONR¹³R¹⁴, and NR¹⁵SO₂R¹⁶, R¹⁵, R¹⁶, R²¹ and R²², each ofwhich when present, is independently selected from the group ofhydrogen, lower alkyl, and aralkyl, R¹³, R¹⁴, R²³ and R²⁴, each ofwhich, when present is independently selected from the group consistingof hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl,heterocyclylalkyl and aralkyl, R³⁴, when present, is selected form thegroup consisting of alkyl, aralkyl, COR³⁵, and SO₂R³⁵, R³⁵ when present,is selected form the group consisting of alkyl, aryl, and aralkyl, andR¹, R², R¹³, R¹⁴, R¹⁵, R¹⁶, R²³, R²⁴, R³⁴ and R³⁵, when present, may beeither unsubstituted or substituted with one or more substituentsselected from the group consisting of alkyl, aryl, aralkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl,hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl,alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino,arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl),—NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl), —NHSO₂(aralkyl),alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino), and—OCO(dialkylamino), with the proviso that when M² is CO, then M⁶ isNR³⁴(CH₂)_(q), wherein q is not 0. In other embodiments, the chemicalcompounds are inactive ingredients or excipients. In other embodiments,the effective amount in carrier is greater than 1 fM and less than 100nM. In other embodiments, the effective amount in carrier is greaterthan 1 fM and less than 50 nM. In other embodiments, the effectiveamount in carrier is greater than 1 fM and less than 25 nM.

The concentration of the disclosed compounds in the ex vivo treatmentmedia alone or in combination with therapeutic cells in a suitable mediato infuse a human or lower animal may be between 1 fM (1 femto molar or1×10⁻¹⁵ M) and less than 10 μM. If desired, the effective concentrationcan be divided into multiple doses for purposes of administration;consequently, single dose compositions may contain such amounts orsubmultiples thereof to make up the daily dose of cell or compound.

In other embodiments, the R¹, R², R³, R¹², R¹⁴, R¹⁶, R²³ and R²⁴ groupswhen present may independently be either unsubstituted or substitutedwith one or more substituents selected from the group consisting ofalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl,heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy,hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl,haloaryl, amino, alkylamino, dialkylamino, arylamino, diarylamino,—NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl),—NHSO₂(alkyl), —NHSO₂(aryl), —NHSO₂(aralkyl), alkoxycarbonyl,alkoxycarbonylalkyl, —OCO(alkylamino), —OCO(dialkylamino), and mixturesor combinations thereof.

In some embodiments, the compound is selected from the group consistingof methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10R)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-1-(1,3-benzodioxol-5-yl)-6-butyl-7-methyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-9-methyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;ethyl(6S,10R)-10-(1,3-benzodioxol-5-yl)-6-butyl-7-methyl-3,8-dioxo-1-(-2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(10S)-10-(1,3-benzodioxol-5-yl)-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-2-methyl-3,8-dioxo-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,-9-triazadodecan-12-oate;(2S)-2-{[(1,3-benzodioxol-5-ylmethyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate;methyl(6S,10S)-6-butyl-3,8-dioxo-10-phenyl-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-({[(1S)-1-(1,3-benzodioxol-5-yl)-3-hydroxypropyl]carbamoyl}amino)hexyl-bis(2-thienylmethyl)carbamate;(2S)-2-[(benzylcarbamoyl)amino]hexyl-bis(2-thienylmethyl)carbamate;(2S)-2-[(morpholin-4-ylcarbonyl)amino]hexyl-bis(2-thienylmethyl)carbamate;(2S)-2-{[(3-methoxypropyl)carbamoyl]amino}hexyl-bis(2-thienylmethyl)carbamate;(2S)-2-{[(2-methoxyethyl)carbamoyl]amino}hexyl-bis(2-thienylmethyl)carbamate;tert-butyl[(2S)-1-{[bis(2-thienylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;(2S)-2-[(tert-butylcarbamoyl)amino]hexyl-bis(2-thienylmethyl)carbamate;(2S)-2-[(isopropylcarbamoyl)amino]hexyl-bis(2-thienylmethyl)carbamate;(2S)-2-[(methylcarbamoyl)amino]hexyl-bis(2-thienylmethyl)carbamate;tert-butyl[(2R)-1-{[bis(2-thienylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;benzyl{(5S)-6-{[bis(2-thienylmethyl)carbamoyl]oxy}-5-[(tert-butoxycarbonyl)amino]hexyl}carbamate;methyl(9S,13S)-13-(1,3-benzodioxol-5-yl)-9-({[bis(2-thienylmethyl)carbamoyl]oxy}methyl)-3,11-dioxo-1-phenyl-2-oxa-4,10,12-triazapentadecan-15-oate;(2S)-2-acetamidohexylbis(2-thienylmethyl)carbamate;methyl(3R)-3-(1,3-benzodioxol-5-yl)-3-{[(2S)-2-{[bis(2-thienylmethyl)carbamoyl]amino}hexanoyl]amino}propanoate;methyl(3R)-3-(1,3-benzodioxol-5-yl)-3-{[(2R)-2-{[bis(2-thienylmethyl)carbamoyl]amino}hexanoyl]amino}propanoate;methyl(3S)-3-(1,3-benzodioxol-5-yl)-3-{[(2R)-2-{[bis(2-thienylmethyl)carbamoyl]amino}hexanoyl]amino}propanoate;methyl(6R,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6R,10R)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(2S)-2-{[bis(2-thienylmethyl)-carbamoyl]amino}hexanoate;methyl(2R)-2-{[bis(2-thienylmethyl)carbamoyl]amino}hexanoate;3-[(2S)-1-hydroxyhexan-2-yl]-1,1-bis(2-thienylmethyl)urea;3-[(2R)-1-hydroxyhexan-2-yl]-1,1-bis(2-thienylmethyl)urea;methyl(2S)-6-{[(benzyloxy)carbonyl]amino}-2-{[bis(2-thienylmethyl)carbamoyl]amino}hexanoate;methyl{[bis(2-thienylmethyl)carbamoyl](methyl)amino}acetate;methyl{[bis(2-thienylmethyl)carbamoyl]amino}acetate;methyl{[bis(2-thienylmethyl)carbamoyl](butyl)amino}acetate;3-(3-hydroxypropyl)-1,1-bis(2-thienylmethyl)urea;methyl(2R)-{[bis(2-thienylmethyl)carbamoyl]amino}(phenyl)acetate;tert-butyl{[bis(2-thienylmethyl)carbamoyl]amino}acetate;tert-butyl{[bis(2-thienylmethyl)carbamoyl](butyl)amino}acetate;benzyl{(5S)-6-{[bis(4-methoxybenzyl)carbamoyl]oxy}-5-[(tert-butoxycarbonyl)amino]hexyl}carbamate;tert-butyl[(2S)-1-{[bis(4-methoxybenzyl)carbamoyl]oxy}hexan-2-yl]carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-2-(4-methoxybenzyl)-1-(4-methoxyphenyl)-3,8-dioxo-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-({[(1S)-1-(1,3-benzodioxol-5-yl)-3-hydroxypropyl]carbamoyl}amino)hexylbis(4-methoxybenzyl)carbamate;(2S)-2-[(tert-butoxycarbonyl)amino]hexyl dibenzylcarbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-2-benzyl-6-butyl-3,8-dioxo-1-phenyl-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(4-methylbenzyl)carbamoyl]oxy}hexan-2-yl]carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-2-(4-methylbenzyl)-1-(4-methylphenyl)-3,8-dioxo-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(4-chlorobenzyl)carbamoyl]oxy}hexan-2-yl]carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-2-(4-chlorobenzyl)-1-(4-chlorophenyl)-3,8-dioxo-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-[(tert-butoxycarbonyl)amino]hexyl(4-bromobenzyl)(2-thienylmethyl)carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-2-(4-bromobenzyl)-6-but-yl-3,8-dioxo-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-2-(4-azidoobenzyl)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-[(tert-butoxycarbonyl)amino]hexylphenyl(2-thienylmethyl)carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-2-phenyl-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(3-thienylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(3-thienyl)-2-(3-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-{[butyl(2-thienylmethyl)carbamoyl]oxy}hexyl]carbamate;(2S)-2-[(tert-butoxycarbonyl)amino]hexylbutyl(2-thienylmethyl)carbamate;methyl(3S,7S)-3-(1,3-benzodioxol-5-yl)-7-butyl-5,10-dioxo-[1-(2-thienylmethyl)-9-oxa-4,6,11-triazapentadecan-1-oate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-{[(2-methoxyethyl)(2-thienyl-methyl)carbamoyl]oxy}hexyl]carbamate;(2S)-2-[(tert-butoxycarbonyl)amino]hexyl(2-methoxyethyl)(2-thienylmethyl)carbamate;methyl(9S,13S)-13-(1,3-benzodioxol-5-yl)-9-butyl-6,11-dioxo-5-(-2-thienylmethyl)-2,7-dioxa-5,10,12-triazapentadecan-15-oate; (2S)-2-[({3-[(methylsulfonyl)amino]benzyl}carbamoyl)amino]hexyl(2-methoxyethyl)(2-thienylmethyl)carbamate;(2S)-2-{[(4-bromobenzyl)carbamoyl]amino}hexyl-bis(2-thienylmethyl)carbamate;(2S)-2-{[(4-azidobenzyl)carbamoyl]amino}hexyl-bis(2-thienylmethyl)carbamate;tert-butyl[(2S)-1-{[bis(2-thienylmethyl)carbamoyl]thio}hexan-2-yl]carbamate;methyl(6S,1S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-thia-2,7,9-triazadodecan-12-oate;and mixtures or combinations thereof.

In some embodiments, a chemical compound is provided having the generalformula (I), where R¹ is aryl or aralkyl, R² is alkyl, aryl or aralkyl,M¹ is CH₂, M² is CO, M³ is absent, M⁴ is absent or is CH₂, M⁵ is(CR¹¹R¹²), M⁶ is (CH₂)_(q), wherein q is an integer of 0 to 6, R¹¹ ishydrogen, and R¹² is selected from the group consisting of hydrogen,NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴, NR²¹COOR²⁴, OCOR²⁴, OR²⁴, SCOR²⁴,SR²⁴, N₃, CN, and O(CH₂CH₂O)_(s)R²⁴, wherein s is an integer of 1 to 6,R²¹ and R²² when present are independently selected from the groupconsisting of hydrogen, lower alkyl, or aralkyl, R²³ when present isselected from the group consisting of hydroxyalkyl, alkoxyalkyl, alkyl,aryl, aralkyl, and alkoxycarbonylalkyl, R²⁴ when present is selectedfrom the group consisting of alkyl, aryl, aralkyl, heterocyclyl,cycloalkyl, cycloalkylalkyl and heterocyclylalkyl, provided that when M³and M⁴ are absent, R¹² is not of the formula:

where A is selected from the group consisting of —O—, —S—, and —NR²⁶—, Eis selected from the group consisting of —CH₂—, —O—, —S—, and —NR²⁷—, Jis selected from the group consisting of —O—, —S—, and —NR²⁸—, T isselected from the group consisting of CO and (CH₂)_(b) wherein b is aninteger of zero to three, L is selected from the group consisting of—(CH₂)—, —O—, —S—, and —NR²⁹—, wherein n is an integer of zero to three,M is selected from the group consisting of CR³⁰R³¹ and (CH₂)_(u)uwherein u is an integer of zero or one, X is selected from the groupconsisting of CO₂B, PO₃H₂, SO₃H, OPO₃H₂, CONHCOR³², CONHSO₂R³³,oxazolyl, tetrazolyl and hydrogen, B, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹,R³² and R³³ are independently selected from the group consisting ofhydrogen, halogen alkyl, alkenyl, alkynyl, alkoxy, alkenoxy, alkynoxy,thioalkoxy, aliphatic acyl, —CF₃, nitro, amino, cyano, N(C₁-C₃alkyl)CO(C₁-C₃ alkyl), C₁-C₃ alkylamino, alkenylamino, alkynylamino,di(C₁-C₃ alkyl)amino, CO₂(C₁-C₃ alkylamino), CONH(C₁-C₃ alkylamino),CH.dbd.NOH, PO₃H₂, OPO₃H₂, CON(C₁-C₃ alkyl)₂, haloalkyl, alkoxycarbonyl,alkoxyalkoxy, carboxaldehyde, carboxamide, cycloalkyl, cycloalkenyl,cycloalkynyl, aryl, aroyl, aryloxy, arylamino, biaryl, thioaryl,heterocyclyl, heterocycloyl, alkylaryl, aralkenyl, aralkyl,alkylheterocyclyc, heterocyclycalkyl, sulfonyl, sulfonamide, carbamate,aryloxyalkyl, carboxyl and CONH(benzyl), wherein B, X, R²⁵, R²⁶, R²⁷,R²⁸, R²⁹, R³⁰, R³¹ and R³² are unsubstituted or substituted with atleast one electron donating or electron withdrawing group, R³ isselected from the group of hydrogen, NR¹⁵COOR¹⁶, NR¹⁵COR¹⁶,NR¹⁵CONR¹³R¹⁴, NR¹⁵SO₂R¹⁶, OCOR¹⁶, COOR¹⁶, alkyl, SR¹⁶, heterocyclyl,hydroxyl, hydroxyalkyl, guanadino and aryl, wherein R¹³ and R¹⁵ whenpresent are independently hydrogen, lower alkyl, or aralkyl, R¹⁴ and R¹⁶when present are independently selected from the group consisting ofhydrogen, alkyl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl andheterocyclylalkyl provided that when R³ is hydrogen, alkyl or aryl, R¹²is not hydrogen, and provided that when R¹ is phenyl, R³ isbenzyloxycarbonylamino, and R¹² is hydrogen, R² is not 2-methoxybenzyl,and R¹, R², R³, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R²¹, R²², R²³ and R²⁴ whenpresent may independently be either unsubstituted or substituted withone or more substituents selected from the group consisting of alkyl,aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl,heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy, azido,hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl,haloaryl, haloalkoxy, amino, alkylamino, dialkylamino, arylamino,diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl),—NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl), —NHSO₂(aralkyl),alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino),—OCO(dialkylamino), or mixtures and combinations thereof.

In some embodiments, a compound is selected from the group consisting of(2R)-2-({[(1S)-1-(1,3-benzodioxol-5-yl)-3-hydroxypropyl]carbamoyl}amino)-N,N-bis(2-thienylmethyl)hexanamide;methyl(3S)-3-(1,3-benzodioxol-5-yl)-3-[({3-[bis(2-thienylmethyl)amino]-3-oxopropyl}carbamoyl)amino]propanoate;(2S)-2-[(tert-butylcarbamoyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;tert-butyl{(2S)-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-yl}carbamate;benzyl{(5S)-6-[bis(2-thienylmethyl)amino]-5-[(tert-butoxycarbonyl)(methyl)amino]-6-oxohexyl}carbamate;benzyl{(5S)-6-[bis(2-thienylmethyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;benzyl{(5R)-6-[bis(2-thienylmethyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;tert-butyl{(2R)-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-yl}carbamate;(2S)-2-acetamido-N,N-bis(2-thienylmethyl)hexanamide;benzyl{(5S)-5-acetamido-6-[bis(2-thienylmethyl)amino]-6-oxohexyl}carbamate;(2R)-2-acetamido-N,N-bis(2-thienylmethyl)hexanamide;benzyl{(5S)-5-(benzoylamino)-6-[bis(2-thienylmethyl)amino]-6-oxohexyl}carbamate;(2S)-2-[(phenylsulfonyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;(2S)-2-[methyl(phenylsulfonyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;2-[(phenylsulfonyl)amino]-N,N-bis(2-thienylmethyl)acetamide;2-[methyl(phenylsulfonyl)amino]-N,N-bis(2-thienylmethyl)acetamide;(2S)-2-[(methylsulfonyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;(2S)-2-({[3-(4-methoxyphenoxy)propyl]sulfonyl}amino)-N,N-bis(2-thienylmethyl)hexanamide;benzyl{(5S)-6-[bis(2-thienylmethyl)amino]-6-oxo-5-[(2-thienylsulfonyl)amino]hexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(3-methoxybenzyl)(2-thienyl-methyl)amino]-6-oxohexyl}carbamate;benzyl{(5S)-6-[bis(3-methoxybenzyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;benzyl{(5R)-5-[(tert-butoxycarbonyl)amino]-6-[(3-methoxybenzyl)(2-thienyl-methyl)amino]-6-oxohexyl}carbamate;benzyl{(5R)-6-[bis(3-methoxybenzyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-oxo-6-{[2-(2-thienyl)ethyl](-2-thienylmethyl)amino}hexyl]carbamate;benzyl[(5R)-5-[(tert-butoxycarbonyl)amino]-6-oxo-6-{[2-(2-thienyl)ethyl](-2-thienylmethyl)amino}hexyl]carbamate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-(dibenzylamino)-6-oxohexyl]carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(4-nitrobenzyl)(2-thienylmethyl)amino]-6-oxohexyl}carbamate;benzyl{(5R)-5-[(tert-butoxycarbonyl) amino]-6-[(4-nitrobenzyl)(2-thienylmethyl)amino]-6-oxohexyl}carbamate;tert-butyl[(2R)-1-[(4-aminobenzyl)(2-thienylmethyl)amino]-6-{[(benzyloxy)-carbonyl]amino}-1-oxohexan-2-yl]carbamate;tert-butyl[(2S)-1-[(4-aminobenzyl)(2-thienylmethyl)amino]-6-{[(benzyloxy)-carbonyl]amino}-1-oxohexan-2-yl]carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[methyl(2-thienylmethyl)amino]-6-oxohexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[butyl(2-thienylmethyl)amino]-6-oxohexyl}carbamate;benzyl{(5S)-6-[bis(4-methoxybenzyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-oxo-6-[(pyridin-4-ylmethyl)(-2-thienylmethyl)amino]hexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-oxo-6-[(pyridin-3-ylmethyl)(-2-thienylmethyl)amino]hexyl}carbamate;benzyl{(5S)-6-[bis(pyridin-4-ylmethyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;tert-butyl{(2S)-1-[bis(2-thienylmethyl)amino]-1-oxo-6-[(2-thienylsulfonyl)amino]hexan-2-yl}carbamate;tert-butyl{(2S)-6-acetamido-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-yl}carbamate;tert-butyl{(2S)-1-[bis(2-thienylmethyl)amino]-1-oxo-6-[(trifluoroacetyl)amino]hexan-2-yl}carbamate;tert-butyl{(2S)-1-[bis(2-thienylmethyl)amino]-6-[(methylsulfonyl)amino]-1-oxohexan-2-yl}carbamate;tert-butyl{(2S)-1-[bis(2-thienylmethyl)amino]-1-oxo-6-[(2-thienylcarbonyl)amino]hexan-2-yl}carbamate;tert-butyl{(2S)-1-[bis(2-thienylmethyl)amino]-1-oxo-6-[(phenylsulfonyl)amino]hexan-2-yl}carbamate;tert-butyl{(2S)-1-[bis(2-thienylmethyl)amino]-1-oxo-6-[(pyridin-3-ylcarbonyl)amino]hexan-2-yl}carbamate;tert-butyl{(2S)-1-[bis(2-thienylmethyl)amino]-1-oxo-6-[(2-thienylacetyl)amino]hexan-2-yl}carbamate;tert-butyl{(2S)-1-[bis(2-thienylmethyl)amino]-6-hydroxy-1-oxohexan-2-yl}carbamate;tert-butyl[(2S)-1-[bis(2-thienylmethyl)amino]-1-oxo-6-{[(trifluoromethyl)sulfonyl]amino}hexan-2-yl]carbamate;tert-butyl{(2S)-6-[(benzylsulfonyl)amino]-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-yl}carbamate;tert-butyl{(2S)-6-[benzyl(trifluoroacetyl)amino]-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-yl}carbamate;tert-butyl[(1R)-2-[bis(2-thienylmethyl)amino]-1-(4-hydroxyphenyl)-2-oxoet-hyl]carbamate;methyl(4S)-5-[bis(2-thienylmethyl)amino]-4-[(tert-butoxycarbonyl)amino]-5-oxopentanoate;benzyl{(3S)-4-[bis(thiophen-2-ylmethyl)amino]-3-[(tert-butoxycarbonyl)amino]-4-oxobutyl}carbamate;benzyl{(4S)-5-[bis(2-thienylmethyl)amino]-4-[(tert-butoxycarbonyl)amino]-5-oxopentyl}carbamate;tert-butyl{2-[bis(2-thienylmethyl)amino]-2-oxoethyl}carbamate;tert-butyl{2-[bis(2-thienylmethyl)amino]-2-oxoethyl}methylcarbamate;N,N-bis(2-thienylmethyl)-6-[(2-thienylsulfonyl)amino]hexanamide;N-{6-[bis(2-thienylmethyl)amino]-6-oxohexyl}thiophene-2-carboxamide;N-{6-[bis(2-thienylmethyl)amino]-6-oxohexyl}-N-(2-thienylmethyl)thiophene-2-carboxamide;N-benzyl-N-{6-[bis(2-thienylmethyl)amino]-6-oxohexyl}thiophene-2-carboxamide;6-[benzyl(2-thienylsulfonyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;6-[methyl(2-thienylsulfonyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;6-[(benzylsulfonyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;6-[(2-thienylacetyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;N-{6-[bis(2-thienylmethyl)amino]-6-oxohexyl}-N-(3-methoxybenzyl)thiophene-1-2-carboxamide;6-[(3-methoxybenzyl)(2-thienylsulfonyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;6-[(benzylsulfonyl)(3-methoxybenzyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;benzyl{6-[bis(2-thienylmethyl)amino]-6-oxohexyl}carbamate;tert-butyl{6-[bis(thiophen-2-ylmethyl)amino]-6-oxohexyl}carbamate;tert-butyl[(2S)-1-[bis(2-thienylmethyl)amino]-3-(4-hydroxyphenyl)-1-oxopr-opan-2-yl]carbamate;Methyl(5S)-6-[bis(2-thienylmethyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexanoate;(2S)-2-[acetyl(methyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;benzyl{(5S)-5-[acetyl(methyl)amino]-6-[bis(2-thienylmethyl)amino]-6-oxohexyl}carbamate;(2S)-6-{[(benzyloxy)carbonyl]amino}-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-ylacetate;tert-butyl{(2S)-6-[benzyl(2-thienylsulfonyl)amino]-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-yl}carbamate;benzyl{(5S)-6-{bis[4-(trifluoromethoxy)benzyl]amino}-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-oxo-6-{(2-thienylmethyl)[2-(trifluoromethyl)benzyl]amino}hexyl]carbamate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-oxo-6-{(2-thienylmethyl)[2-(trifluoromethoxy) benzyl]amino}hexyl]carbamate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-{[2-(difluoromethoxy)benzyl]-(2-thienylmethyl)amino}-6-oxohexyl]carbamate;tert-butyl{6-[bis(4-methoxybenzyl)amino]-6-oxohexyl}carbamate;N-{6-[bis(4-methoxybenzyl)amino]-6-oxohexyl}-4-methoxybenzamide;N-{6-[bis(4-methoxybenzyl)amino]-6-oxohexyl}-4-methoxy-N-(4-methoxybenzyl-)benzamide;N-{6-[bis(2-thienylmethyl)amino]-6-oxohexyl}-N-methylthiophene-2-carboxamide;6-[(3-methoxybenzyl)(2-thienylacetyl)amino]-N,N-bis(2-thienylmethyl)h-exanamide;tert-butyl{4-[bis(2-thienylmethyl)amino]-4-oxobutyl}carbamate;methyl(3S)-3-(1,3-benzodioxol-5-yl)-3-[({4-[bis(2-thienylmethyl)amino]-4-oxobutyl}carbamoyl)amino]propanoate;6-{[(3-chloropropyl)sulfonyl]amino}-N,N-bis (4-methoxybenzyl)hexanamide;6-(1,1-dioxido-1,2-thiazolidin-2-yl)-N,N-bis(4-methoxybenzyl)hexanamide;N,N-bis(4-methoxybenzyl)-6-({[2-(morpholin-4-yl)ethyl]sulfonyl}amino)hexanamide;3-{[bis(2-thienylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)propanamide;tert-butyl{3-[bis(2-thienylmethyl)amino]-3-oxopropyl}butylcarbamate;3-{[bis(2-thienylmethyl)carbamoyl](butyl)amino}-N,N-bis(2-thienylmethyl)propanamide;3-{butyl[(2-thienylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)propanamide;4-(1,1-dioxido-1,2-thiazolidin-2-yl)-N,N-bis(2-thienylmethyl)butanamide;N,N-bis(2-thienylmethyl)-3-{[(2-thienylmethyl)carbamoyl]amino}propanamide;benzyl{(5S)-6-[bis(2-thienylmethyl)amino]-5-hydroxy-6-oxohexyl}carbamate;benzyl{(5S)-6-[bis(2-thienylmethyl)amino]-5-cyano-6-oxohexyl}carbamate;benzyl{(5R)-5-azido-6-[bis(2-thienylmethyl)amino]-6-oxohexyl}carbamate;S-{(2R)-6-{[(benzyloxy)carbonyl]amino}-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-yl}ethanethioate;tert-butyl[(2S)-1-[bis(2-thienylmethyl)amino]-6-({[(4-bromobenzyl)oxy]carbonyl}amino)-1-oxohexan-2-yl]carbamate;4-azidobenzyl{(5S)-6-[bis(2-thienylmethyl)amino]-5-[(tert-butoxycarbonyl)-amino]-6-oxohexyl}carbamate;benzyl{(5S)-6-[(4-bromobenzyl)(2-thienylmethyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;tert-butyl[(2S)-1-[(4-azidobenzyl)(2-thienylmethyl)amino]-6-{[(benzyloxy)-carbonyl]amino}-1-oxohexan-2-yl]carbamate;tert-butyl{(2S)-1-[(4-bromobenzyl)(2-thienylmethyl)amino]-1-oxohexan-2-yl}carbamate;benzyl{(5S)-6-[bis(3-thienylmethyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(cyclopropylmethyl)(2-thienylmethyl)amino]-6-oxohexyl}carbamate,and mixtures or combinations thereof.

In some embodiments, a chemical compound is provided having the generalformula (I), where R¹ is alkyl, aryl or aralkyl, R² is selected from thegroup consisting of aralkyl and alkyl, provided that when R¹ is alkyl,R² is aralkyl, M¹ is CO or SO₂, provided that when M¹ is SO₂ and R¹ isphenyl, 4-methylphenyl or 2,4,6-trimethylphenyl, R² is not alkyl,2-phenethyl, benzyl, or 2-methoxy-2-oxoethyl, and when M′ is CO and R′is 2-furyl, 4-pyridyl, or 3,5-dinitrophenyl, R² is not alkyl, benzyl or2-(1H-indol-2-yl)ethyl, M² is absent or CH₂, M³ and M⁴ are absent, M⁵ is(CR¹¹R¹²), R¹¹ is hydrogen, R¹² is selected from the group consisting ofhydrogen, NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴, NR²¹COOR²⁴, CONR²²R²³,COOR²⁴, O(CH₂CH₂O)_(s)R²⁴ hydroxyalkyl and alkoxyalkyl, wherein s is aninteger of 1 to 6, M⁶ is (CH₂)_(q) where q is an integer of 0 to 6, R³is selected from the group consisting of NR¹⁵COOR¹⁶, NR¹⁵COR¹⁶,NR¹⁵CONR¹³R¹⁴, and NR¹⁵SO₂R¹⁶, and R¹³, R²¹ and R²², when present, areindependently selected from the group consisting of hydrogen and loweralkyl, and R¹⁴, R¹⁵, R¹⁶, R²³ and R²⁴, each of which when present, isindependently selected from the group consisting of hydrogen, alkyl,aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl andaralkyl, and R¹, R², R³, R¹², R¹⁴, R¹⁵, R¹⁶, R²³ and R²⁴ when presentmay independently be either unsubstituted or substituted with one ormore substituents selected from the group consisting of alkyl, aryl,aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl,heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl,aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino,alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl),—NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO₂(alkyl),—NHSO₂(aryl), —NHSO₂(aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl,—OCO(alkylamino), —OCO(dialkylamino), and mixtures or combinationsthereof.

In some embodiments, a compound is selected from the group consisting ofmethyl(2S)-6-{[(benzyloxy)carbonyl]amino}-2-[benzyl(2-thienylsulfonyl)amino]hexanoate;methyl(2S)-6-{[(benzyloxy)carbonyl]amino}-2-[benzyl(phenylsulfonyl)amino]hexanoate;methyl(2S)-6-{[(benzyloxy)carbonyl]amino}-2-[(2-thienylcarbonyl-)(2-thienylmethyl)amino]hexanoate;methyl(2S)-6-{[(benzyloxy)carbonyl]amino}-2-[(2-thienylacetyl)(2-thienylmethyl)amino]hexanoate;methyl(2S)-2-[benzyl(isobutylsulfonyl)amino]-6-{[(benzyloxy)carbonyl]amino}hexanoate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(2-thienylmethyl)(2-thienylsulfonyl)amino]hexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(2-thienylacetyl)(2-thienylmethyl)amino]hexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(methylsulfonyl)(2-thienylmethyl)amino]hexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(phenylsulfonyl)(2-thienylmethyl)amino]hexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(2-thienylcarbonyl)(2-thienylmethyl)amino]hexyl}carbamate;N,N′-heptane-1,7-diylbis[N-(2-thienylmethyl)benzamide];N,N′-heptane-1,7-diylbis[N-(2-thienylmethyl) thiophene-2-carboxamide];benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-{[(4-methoxyphenyl)sulfonyl]-(2-thienylmethyl)amino}hexyl]carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(4-methoxybenzoyl)(2-thienylmethyl)amino]hexyl}carbamate;N,N′-hexane-1,6-diylbis[N-(2-thienylmethyl) thiophene-2-carboxamide];N,N′-hexane-1,6-diylbis[N-(3-methoxybenzyl) thiophene-2-carboxamide];tert-butyl{5-[(4-methoxybenzyl)(2-thienylsulfonyl)amino]pentyl}carbamate;N,N′-pentane-1,5-diylbis[N-(3-methoxybenzyl)thiophene-2-sulfonamide];N-(3-methoxybenzyl)-N-{5-[(2-thienylsulfonyl)amino]pentyl}thiophene-2-sulfonamide;tert-butyl{5-[(2-thienylcarbonyl)(2-thienylmethyl)amino]pentyl}carbamate;N-(3-methoxybenzyl)-N-{5-[(2-thienylcarbonyl)amino]pentyl}thiophene-2-carboxamide;N,N′-pentane-1,5-diylbis[N-(3-methoxybenzyl)thiophene-2-carboxamide];and mixtures or combinations thereof.

In some embodiments, a chemical compound is selected having the generalformula (I) where R¹ is aryl or aralkyl, R² is alkyl or aralkyl, M¹ isCH₂, M² is CO, M³ is absent or is O or CH₂, M⁴ is absent or is CH₂, M⁵is absent or is O or (CR¹¹R¹²), R¹¹ is hydrogen, R¹² is selected fromthe group consisting of hydrogen, NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴and NR²¹COOR²⁴, M⁶ is selected from the group consisting of (CH₂)_(q),(CH₂)_(q)—CH═CH—(CH₂)_(r), (CH₂)_(q)-arylene-(CH₂)_(r) and(CH₂CH₂O)_(q), wherein q and r are independently integers from 0 to 6,R³ is CONR¹³R¹⁴, R²¹ and R²² each of which, when present isindependently selected from the group of hydrogen and lower alkyl, R¹³,R¹⁴, R²³ and R²⁴, each of which, when present is independently selectedfrom the group consisting of hydrogen, alkyl, aryl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, and R¹,R², R¹³, R¹⁴, R²³ and R²⁴ when present may be either unsubstituted orsubstituted with one or more substituents selected from the groupconsisting of alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl,heterocyclyl, heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy,azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl, halo,haloalkyl, haloaryl, amino, alkylamino, dialkylamino, arylamino,diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl),—NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl), —NHSO₂(aralkyl),alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino),—OCO(dialkylamino), and mixtures or combinations thereof.

In some embodiments, a compound is selected from the group consisting ofN,N,N′,N′-tetrakis(2-thienylmethyl)pentanediamide;N-(3-methoxybenzyl)-N,N′,N′-tris(2-thienylmethyl)pentanediamide;N,N,N′-tris(2-thienylmethyl)pentanediamide;N′-[2-(2-thienyl)ethyl]-N,N-bis(2-thienylmethyl)pentanediamide;N-[2-(2-thienyl)ethyl]-N,N′,N′-tris (2-thienylmethyl)pentanediamide;N,N-bis(pyridin-4-ylmethyl)-N′,N′-bis(2-thienylmethyl) pentanediamide;N,N-bis(pyridin-3-ylmethyl)-N′,N′-bis(2-thienylmethyl)pentanediamide;N,N-bis(3-methoxybenzyl)-N′,N′-bis(2-thienylmethyl)pentanediamide;N,N,N′,N′-tetrakis (4-methoxybenzyl)pentanediamide;N,N,N′,N′-tetrakis(2-thienylmethyl)hexanediamide;N,N,N′,N′-tetrakis(4-methoxybenzyl)hexanediamide;N,N,N′,N′-tetrakis(3-methoxybenzyl) hexanediamide;N,N,N′,N′-tetrakis(2-thienylmethyl)heptanediamide;2,2′-(1,3-phenylene)bis[N,N-bis (2-thienylmethyl)acetamide];N,N,N′,N′-tetrakis(4-methoxybenzyl)heptanediamide;N,N,N′,N′-tetrakis(2-thienylmethyl)octanediamide;(3E)-N,N,N′,N′-tetrakis(2-thienylmethyl) hex-3-enediamide;2,2′-oxybis[N,N-bis(2-thienylmethyl)acetamide];3-oxo-1-(2-thienyl)-2-(2-thienylmethyl)-4,7,10-trioxa-2-azadodecan-12-ylbis(2-thienylmethyl)carbamate;N,N,N′,N′-tetrakis(4-methoxybenzyl)succinamideethane-1,2-diylbis[bis(2-thienylmethyl)carbamate];N,N,N′,N′-tetrakis(4-methoxybenzyl)octanediamide;N,N,N′,N′-tetrakis(2-thienylmethyl)pyridine-3,5-dicarboxamide;N,N,N′,N′-tetrakis(2-thienylmethyl) pyridine-2,6-dicarboxamide;N,N,N′,N′-tetrakis(2-thienylmethyl)pyridine-2,4-dicarboxamide;2,2′-(1,4-phenylene)bis[N,N-bis(2-thienylmethyl)acetamide];8-{2-[bis(2-thienylmethyl)amino]-2-oxoethoxy}-N,N-bis(2-thienylmethyl)quinoline-2-carboxamide;N,N′-bis(4-methoxybenzyl)-N,N′-bis(2-thienylmethyl)hexanediamide;tert-butyl{(2S)-1,6-bis[bis(2-thienylmethyl)amino]-1,6-dioxohexan-2-yl}carbamate;and mixtures or combinations thereof.

In some embodiments a chemical compound is provided having the generalformula (I), where R¹ is aryl or aralkyl, R² is alkyl or aralkyl, M¹ isCH₂, M² is SO₂ or CO, M³ is absent or is CH₂, M⁴ is absent or is CH₂, M⁵is absent or is (CR¹¹R¹²), R¹¹, when present, is hydrogen, R¹², whenpresent, is selected from the group consisting of hydrogen, alkyl,NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴ and NR²¹COOR²⁴, M⁶ is (CH₂)_(q), orNR³⁴(CH₂)_(q), wherein q is an integer from 0 to 6, R³ is selected fromthe group consisting of CONR¹³R¹⁴, SO₂NR¹³R¹⁴, NR¹⁵COOR¹⁶, NR¹⁵COR¹⁶,NR¹⁵CONR¹³R¹⁴, and NR¹⁵SO₂R¹⁶, R¹⁵, R¹⁶, R²¹ and R²², each of which whenpresent, is independently selected from the group of hydrogen, loweralkyl, and aralkyl, R¹³, R¹⁴, R²³ and R²⁴, each of which, when presentis independently selected from the group consisting of hydrogen, alkyl,aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl andaralkyl, R³⁴, when present, is selected form the group consisting ofalkyl, aralkyl, COR³⁵, and SO₂R³⁵, where R³⁵ when present, is selectedform the group consisting of alkyl, aryl, and aralkyl, and R¹, R², R¹³,R¹⁴, R¹⁵, R¹⁶, R²³, R²⁴, R³⁴ and R³⁵, when present, may be eitherunsubstituted or substituted with one or more substituents selected fromthe group consisting of alkyl, aryl, aralkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl,hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl,alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino,arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl),—NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl), —NHSO₂(aralkyl),alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino), and—OCO(dialkylamino), with the proviso that when M² is CO, then M⁶ isNR³⁴(CH₂)_(q), wherein q is not 0.

In some embodiments, a compound is selected from the group consisting ofN-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}-N-(2-thienylmethyl)thiophene-2-sulfonamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}-N-(2-thienylmethyl)thiophene-2-carboxamide;2-{butyl[(2-thienylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)ethanesulfonamide;2-{[bis(2-thienylmethyl)carbamoyl](butyl)amino}-N,N-bis(2-thienylmethyl)ethanesulfonamide;N-{3-[bis(2-thienylmethyl)sulfamoyl]propyl}-N-(2-thienylmethyl)thiophene-2-sulfonamide;2-[(methylsulfonyl)(2-thienylmethyl)amino]-N,N-bis(2-thienylmethyl)ethanesulfonamide;2-{[bis(2-thienylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)ethanesulfonamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}thiophene-2-sulfonamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}-2-(2-thienyl)acetamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}thiophene-2-carboxamide;N,N-bis(2-thienylmethyl)-2-{[(2-thienylmethyl)carbamoyl]amino}ethanesulfonamide;2-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)acetamide;3-[{2-[bis(2-thienylmethyl)amino]-2-oxoethyl}(butyl)amino]-N,N-bis(2-thienylmethyl)propanamide;2-[{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}(methyl)amino]-N,N-bis(2-thienylmethyl)acetamide;2-[{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}(butyl)amino]-N,N-bis(2-thienylmethyl)acetamide;3-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)propanamide;3-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(4-methoxybenzyl)propanamide;3-({2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)propanamide;3-[{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}(methyl)amino]-N,N-bis(2-thienylmethyl)propanamide;3-[{2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}(methyl)amino]-N,N-bis(2-thienylmethyl)propanamide;(2S)-2-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)hexanamide;(2S)-2-({2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)hexanamide;2-(acetyl{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)acetamide;2-(acetyl{2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienyl-methyl)acetamide;and mixtures or combinations thereof.

In some embodiments, a compound is selected from the group consisting oftert-butyl[(2S)-1-{[bis(cyclopropylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;(2S)-2-[(tert-butoxycarbonyl)amino]hexyldiisobutylcarbamate;methyl(8S,12S)-12-(1,3-benzodioxol-5-yl)-8-butyl-4-isobutyl-2-methyl-5,10-dioxo-6-oxa-4,9,11-triazatetradecan-14-oate;benzyl{(5S)-6-[bis(cyclopropylmethyl)amino]-5-[(tert-butoxycarbonyl)amino-]-6-oxohexyl}carbamate;and mixtures or combinations thereof.

In accordance with certain embodiments, a pharmaceutical composition isprovided comprising an above-described compound or a pharmaceuticallyacceptable salt thereof; and a pharmaceutically acceptable carrier.

In accordance with certain embodiments, a method of treatingintegrin-expressing cells is provided. The integrin may be one or moreof α4β1, α5β1, α4β7, αvβ3 and αLβ2, for example. In some embodiments,the method of treating integrin-expressing cells comprises contacting atleast one integrin-expressing cell in vitro with an agonist of saidintegrin, wherein said agonist is a compound having the general formula(I), where R¹ and R² are independently selected from the groupconsisting of alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl,haloalkyl, heterocylcyl and heterocyclylalkyl, one of M¹ and M² is CO orSO₂ and the other is (CR⁴R⁵)_(l), provided that when M² is CO, M³ is O,S, NR⁶ or (CR⁷R⁸)_(m), and provided that when M² is SO₂ or (CR⁴R⁵)_(l),M³ is (CR⁷R⁸)_(m), M⁴ is absent or (CR⁹R¹⁰)_(n), M⁵ is absent or is O or(CR¹¹R¹²), M⁶ is absent or is selected from the group consisting of(CH₂)_(q), (CH₂)_(q)—CH═CH—(CH₂)_(r), (CH₂)_(q)-arylene-(CH₂)_(r),(CH₂CH₂O)_(q), and NR³⁴(CH₂)_(q), and R³ is selected from the groupconsisting of hydrogen, OH, OR¹⁶, CONR¹³R¹⁴, NR¹⁵COOR¹⁶, NR¹⁵COR¹⁶,NR¹⁵CONR¹³R¹⁴, NR¹⁵SO₂R¹⁶, OCOR¹⁶, COOR¹⁶, alkyl, aryl, aralkyl, SR¹⁶,heterocyclyl, hydroxyalkyl and guanadino, R³⁴, when present, is selectedform the group consisting of alkyl, aralkyl, COR³⁵, and SO₂R³, R³⁵, whenpresent, is selected form the group consisting of alkyl, aryl, andaralkyl, and R¹², when present, is selected from the group consisting ofhydrogen, alkyl, OH, N₃, CN, NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹COOR²⁴,NR²¹SO₂R²⁴, CONR²²R²³, COOR²⁴, OCOR²⁴, OR²⁴, SCOR²⁴, SR²⁴, azido, CN,and O(CH₂CH₂O)_(s)R²⁴, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹⁵, and R²¹,each of which when present, is independently selected from the groupconsisting of hydrogen, lower alkyl and aralkyl, R¹³, R¹⁴, R¹⁶, R²², R²³and R²⁴, each of which when present, is independently selected from thegroup consisting of alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl,haloalkyl, heterocyclyl, heterocyclylalkyl, alkoxycarbonyl,alkoxycarbonylalkyl, 1, m, n and p are independently integers from 0 to1, q, r and s are independently integers from 0 to 6, R¹, R², R³, R⁴,R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R²¹, R²², R²³,R²⁴, R³⁴ and R³⁵, each of which when present, is independently eitherunsubstituted or substituted with one or more substituents selected fromthe group consisting of alkyl, aryl, aralkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl,hydroxyl, alkoxy, haloalkoxy, azido, hydroxyalkyl, aryloxy, hydroxyaryl,alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino,arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(haloalkyl),—NHSO₂(alkyl), —NHSO₂(aryl), alkoxycarbonyl, alkoxycarbonylalkyl,—OCO(alkylamino), —OCO(dialkylamino), and mixtures or combinationsthereof.

In accordance with certain embodiments, a method of enhancing binding ofcells to an integrin-binding ligand is provided, wherein the methodcomprises treating integrin-expressing cells in vitro with an agonist ofintegrin described above, wherein said integrin is selected from thegroup consisting of α4β1, α5β1, α4β7, αvβ3 and αLβ2; and contacting thetreated cells with an integrin-binding ligand.

In some embodiments, the agonist of integrin utilized in an abovedescribed method is a compound selected from the group consisting ofmethyl(3R)-3-(1,3-benzodioxol-5-yl)-3-[({(2R)-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-yl}carbamoyl)amino]propanoate;methyl(3S)-3-(1,3-benzodioxol-5-yl)-3-[({(2R)-6-{[(benzyloxy)carbonyl]amino}-1-[bis(thiophen-2-ylmethyl)amino]-1-oxohexan-2-yl}carbamoyl)amino]propanoate;methyl(3S)-3-(1,3-benzodioxol-5-yl)-3-[({(2R)-1-[bis(thiophen-2-yl-methyl)amino]-1-oxohexan-2-yl}carbamoyl)amino]propanoate;methyl(3R)-3-(1,3-benzodioxol-5-yl)-3-[({(2S)-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-yl}carbamoyl)amino]propanoate;methyl(3S)-3-(1,3-benzodioxol-5-yl)-3-[({2-[bis(2-thienylmethyl)amino]-2-oxoethyl}carbamoyl)amino]propanoate;methyl(3S)-3-(1,3-benzodioxol-5-yl)-3-[({2-[bis(2-thienylmethyl)amino]-2-oxoethyl}carbamoyl)amino]propanoate;methyl(3S)-3-(1,3-benzodioxol-5-yl)-3-{[{2-[bis(2-thienylmethyl)amino]-2-oxoethyl}(methyl)carbamoyl]amino}propanoate;methyl(3R)-3-(1,3-benzodioxol-5-yl)-3-{[{2-[bis(2-thienylmethyl)amino]-2-oxoethyl}(methyl)carbamoyl]amino}propanoate;methyl(3R)-3-(1,3-benzodioxol-5-yl)-3-[({2-[bis(2-thienylmethyl)amino]-2-oxoethyl}carbamoyl)amino]propanoate;methyl(2R)-[({(2S)-1-[bis(thiophen-2-ylmethyl)amino]-1-oxohexan-2-yl}carbamoyl)amino](phenyl)ethanoate;methyl3-[({(2S)-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-yl}carbamoyl)amino]propanoate;(2S)-2-[(isopropylcarbamoyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;(2S)-2-[(methylcarbamoyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;(2S)-2-[(benzylcarbamoyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;(2R)-2-[(benzylcarbamoyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;benzyl{(5S)-5-[(benzylcarbamoyl)amino]-6-[bis(2-thienylmethyl)amino]-6-ox-ohexyl}carbamate;(2S)-2-{[(1,3-benzodioxol-5-ylmethyl)carbamoyl]amino}-N,N-bis(2-thienylme-thyl)hexanamide;benzyl[(5S)-6-[bis(2-thienylmethyl)amino]-6-oxo-5-{[(pyridin-3-ylmethyl)carbamoyl]amino}hexyl]carbamate;(2S)-2-{[(pyridin-3-ylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)hexanamide;(2S)-2-({[(6-methoxypyridin-3-yl)methyl]carbamoyl}amino)-N,N-bis(-2-thienylmethyl)hexanamide;(2S)-2-({[3-(morpholin-4-yl)benzyl]carbamoyl}amino)-N,N-bis(2-thienylmethyl)hexanamide;(2S)-2-{[(4-hydroxybenzyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)hexanamide;(2S)-2-({[4-(dimethylamino)benzyl]carbamoyl}amino)-N,N-bis(2-thienylmethyl)hexanamide;benzyl[(5S)-6-[bis(2-thienylmethyl)amino]-5-({[3-(morpholin-4-yl)benzyl]carbamoyl}amino)-6-oxohexyl]carbamate;benzyl{(5S)-6-[bis(2-thienylmethyl)amino]-5-[({3-[(methylsulfonyl)amino]benzyl}carbamoyl)amino]-6-oxohexyl}carbamate;benzyl{(2S)-6-{[(benzyloxy)carbonyl]amino}-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-yl}carbamate;benzyl{(2S)-1-[bis(2-thienylmethyl)amino]-1-oxohexan-2-yl}carbamate;benzyl{(5S)-6-[bis(2-thienylmethyl)amino]-5-[(ethoxycarbonyl)amino]-6-oxohexyl}carbamate;benzyl[(5S)-6-[bis(2-thienylmethyl)amino]-5-(butyrylamino)-6-oxohexyl]carbamate;benzyl{(5S)-6-[bis(2-thienylmethyl)amino]-6-oxo-5-[(3-phenoxypropanoyl)amino]hexyl}carbamate;and mixtures or combinations thereof.

In other embodiments, an integrin agonist used in a method of enhancingbinding of cells to an integrin-binding ligand is selected from thegroup consisting of compounds having the general formula (I) where R¹ isselected from the group consisting of alkyl, aryl, and aralkyl, R² isselected from the group consisting of alkyl, aryl, aralkyl, alkoxyalkyland hydroxyalkyl, M¹ is CH₂, M² is SO₂; M³, M⁴, M⁵, and M⁶ independentlyare absent or are CH₂; R³ is selected from the group consisting ofalkyl, aryl, aralkyl, aryloxyalkyl, alkoxyalkyl, cycloalkyl andcycloalkylalkyl; R¹, R² and R³ are independently either unsubstituted orsubstituted with one or more substituents selected from the groupconsisting of alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl,heterocyclyl, heterocyclylalkyl, heterocyclylaryl, hydroxyl, alkoxy,hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl,haloaryl, amino, alkylamino, dialkylamino, arylamino, diarylamino,—NHCO(alkyl), —NHCO(aryl), —NHCO(haloalkyl), —NHSO₂(alkyl),—NHSO₂(aryl), alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino),—OCO(dialkylamino), and mixtures or combinations thereof.

In accordance with some further embodiments, an integrin agonist used ina method of enhancing binding of cells to an integrin-binding ligand isselected from the group consisting ofN-bis(2-thienylmethyl)benzenesulfonamide;N,N-bis(2-thienylmethyl)acetamide;1-phenyl-N,N-bis(2-thienylmethyl)methanesulfonamide;2-methyl-N,N-bis(2-thienylmethyl) propane-1-sulfonamide;N-(3-methoxybenzyl)-N-(2-thienylmethyl)benzenesulfonamide;N-(3-methoxybenzyl)-N-(2-thienylmethyl)propane-2-sulfonamide;N-(3-methoxybenzyl)-2-methyl-(2-thienylmethyl)propane-1-sulfonamide;N-(4-hydroxybenzyl)-3-methoxy-N-(2-thienylmethyl) benzenesulfonamide;N-[2-(2-thienyl)ethyl]-N-(2-thienylmethyl)benzenesulfonamide;N,N-dibenzylbenzenesulfonamide;N-(pyridin-3-ylmethyl)-N-(2-thienylmethyl)benzenesulfonamide;N-butyl-N-(2-thienylmethyl)benzenesulfonamide;N-(3-hydroxypropyl)-N-(2-thienylmethyl) benzenesulfonamide;N-(2-methoxyethyl)-N-(2-thienylmethyl)benzenesulfonamide;N-(2-methoxyethyl)-N-(2-thienylmethyl)thiophene-2-sulfonamide;N,N-bis(3-methoxybenzyl)benzenesulfonamide; N,N-bis(4-methoxybenzyl)thiophene-2-sulfonamide;2-chloro-N,N-bis(2-thienylmethyl)benzenesulfonamide;3-chloro-N,N-bis(2-thienylmethyl)benzenesulfonamide;4-chloro-N,N-bis(2-thienylmethyl) benzenesulfonamide;3-methoxy-N,N-bis(2-thienylmethyl)benzenesulfonamide;4-methoxy-N,N-bis(2-thienylmethyl)benzenesulfonamide;N,N-bis(pyridin-4-ylmethyl )benzenesulfonamide;N,N-bis(pyridin-3-ylmethyl)benzenesulfonamide;N-(2-furylmethyl)-N-(2-thienylmethyl)benzenesulfonamide;N,N-bis(2-furylmethyl) benzenesulfonamide;N,N-bis(3-methoxybenzyl)thiophene-2-sulfonamide; methyl3-[bis(3-methoxybenzyl)sulfamoyl]thiophene-2-carboxylate;2-(hydroxymethyl)-N,N-bis (3-methoxybenzyl)thiophene-3-sulfonamide;N,N-bis(4-methoxybenzyl)-3-methylbenzenesulfonamide;N-phenyl-N-(2-thienylmethyl)benzenesulfonamide;N-phenyl-N-(2-thienylmethyl)thiophene-2-sulfonamide;N-(3-methoxybenzyl)-phenylthiophene-2-sulfonamide;N-(3-methoxybenzyl)-N-phenylbenzenesulfonamide;3-(4-methoxyphenoxy)-N,N-bis(2-thienylmethyl)propane-1-sulfonamide;4-methyl-N,N-bis(2-thienylmethyl)benzenesulfonamide;2-methyl-N,N-bis(2-thienylmethyl) benzenesulfonamide;3-methyl-N,N-bis(2-thienylmethyl)benzenesulfonamide; and mixtures orcombinations thereof.

In other embodiments a method of enhancing binding of cells to anintegrin-binding ligand is provided, wherein said agonist of integrin isa compound selected from the group consisting ofmethyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-7-methyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(2-thienylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;(2S)-2-{[(1,3-benzodioxol-5-ylmethyl)carbamoyl]amino}hexyl-bis(2-thienylmethyl)carbamate;methyl(6S,10S)-6-butyl-3,8-dioxo-10-phenyl-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-[(benzylcarbamoyl)amino]hexyl bis(2-thienylmethyl) carbamate;(2S)-2-({[(1S)-1-(1,3-benzodioxol-5-yl)-3-hydroxypropyl]carbamoyl}amino)hexylbis(2-thienylmethyl)carbamate;methyl(6S,10R)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;benzyl{(5S)-6-{[bis(2-thienylmethyl)carbamoyl]oxy}-5-[(tert-butoxycarbonyl)amino]hexyl}carbamate;methyl(9S,13S)-13-(1,3-benzodioxol-5-yl)-9-({[bis(2-thienylmethyl)carbamoyl]oxy}methyl)-3,11-dioxo-1-phenyl-2-oxa-4,10,12-triazapentadecan-15-oate;tert-butyl[(2R)-1-{[bis(2-thienylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;tert-butyl{[bis(2-thienylmethyl)carbamoyl](butyl)amino}acetate;benzyl{(5S)-6-{[bis(4-methoxybenzyl)carbamoyl]oxy}-5-[(tert-butoxycarbonyl)amino]hexyl}carbamate;tert-butyl[(2S)-1-{[bis(4-methoxybenzyl)carbamoyl]oxy}hexan-2-yl]carbamate;methyl(6S,10S)-1-(1,3-benzodioxol-5-yl)-6-butyl-2-(4-methoxybenzyl)-1-(4-methoxyphenyl)-3,8-dioxo-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-({[(1S)-1-(1,3-benzodioxol-5-yl)-3-hydroxypropyl]carbamoyl}amino)hexyl-bis(4-methoxybenzyl)carbamate;(2S)-2-[(tert-butoxycarbonyl)amino]hexyldibenzylcarbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-2-benzyl-6-butyl-3,8-dioxo-1-phe-nyl-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(4-methylbenzyl)carbamoyl]oxy}hexan-2-yl]carbamate-methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-2-(4-methylbenzyl)-1-(4-methylphenyl)-3,8-dioxo-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(4-chlorobenzyl)carbamoyl]oxy}hexan-2-yl]carbamate;methyl(6S,10S)-1-(1,3-benzodioxol-5-yl)-6-butyl-2-(4-chlorobenzyl)-1-(4-chlorophenyl)-3,8-dioxo-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-2-(4-bromobenzyl)-6-butyl-3,8-dioxo-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-2-(4-azidoobenzyl)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-[(tert-butoxycarbonyl)amino]hexylphenyl(2-thienylmethyl)carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-2-phenyl-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(3-thienylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(3-thienyl)-2-(3-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-{[butyl(2-thienylmethyl)carbamoyl]oxy}hexyl]carbamate;(2S)-2-[(tert-butoxycarbonyl)amino]hexylbutyl(2-thienylmethyl)carbamate;methyl(3S,7S)-3-(1,3-benzodioxol-5-yl)-7-butyl-5,10-dioxo-11-(2-thienylmethyl)-9-oxa-4,6,11-triazapentadecan-1-oate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-{[(2-methoxyethyl)(2-thienylmethyl)carbamoyl]oxy}hexyl]carbamate;(2S)-2-{[(4-bromobenzyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate;(2S)-2-{[(4-azidobenzyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate;tert-butyl[(2S)-1-{[bis(2-thienylmethyl)carbamoyl]thio}hexan-2-yl]carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-thia-2,7,9-triazadodecan-12-oate;and mixtures or combinations thereof.

In some embodiments, a method of enhancing binding of cells to anintegrin-binding ligand is provided, wherein an agonist of integrin is acompound selected from the group consisting ofbenzyl{(5R)-5-[(tert-butoxycarbonyl)amino]-6-[(3-methoxybenzyl)(2-thienylmethyl)amino]-6-oxohexyl}carbamate;benzyl{(5R)-6-[bis(3-methoxybenzyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;benzyl{(5S)-6-[bis(4-methoxybenzyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-oxo-6-[(pyridin-3-ylmethyl)(-2-thienylmethyl)amino]hexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-oxo-6-[(pyridin-4-ylmethyl)(2-thienylmethyl)amino]hexyl}carbamate; (2S)-2-[methyl(phenylsulfonyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;(2S)-2-({[3-(4-methoxyphenoxy)propyl]sulfonyl}amino)-N,N-bis(2-thienylmet-hyl)hexanamide;benzyl{(5R)-6-[bis(2-thienylmethyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;benzyl{(5S)-6-[bis(2-thienylmethyl)amino]-5-[(tert-butoxycarbonyl)amino]-6-oxohexyl}carbamate;benzyl{(5S)-6-[bis(2-thienylmethyl)amino]-6-oxo-5-[(2-thienylsulfonyl)amino]hexyl}carbamate;tert-butyl{(2S)-1-[bis(2-thienylmethyl)amino]-1-oxo-6-[(2-thienylsulfonyl)amino]hexan-2-yl}carbamate;6-[methyl(2-thienylsulfonyl)amino]-N,N-bis (2-thienylmethyl)hexanamide;6-[(2-thienylacetyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;benzyl{(4S)-5-[bis(2-thienylmethyl)amino]-4-[(tert-butoxycarbonyl)amino]-5-oxopentyl}carbamate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-oxo-6-{(2-thienylmethyl)[2-(trifluoromethyl)benzyl]amino}hexyl]carbamate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-oxo-6-{(2-thienylmethyl)[2-(trifluoromethoxy)benzyl]amino}hexyl]carbamate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-{[2-(difluoromethoxy)benzyl]-(2-thienylmethyl)amino}-6-oxohexyl]carbamate;tert-butyl{6-[bis(4-methoxybenzyl)amino]-6-oxohexyl}carbamate;N-{6-[bis(4-methoxybenzyl)amino]-6-oxohexyl}-4-methoxy-N-(4-methoxybenzyl)benzamide;N-{6-[bis(2-thienylmethyl)amino]-6-oxohexyl}-N-methylthiophene-2-carboxamide; 6-[(3-methoxybenzyl)(2-thienylacetyl)amino]-N,N-bis(2-thienylmethyl)hexanamide;methyl(3S)-3-(1,3-benzodioxol-5-yl)-3-[({4-[bis(2-thienylmethyl)amino]-4-oxobutyl}carbamoyl)amino]propanoate;6-{[(3-chloropropyl)sulfonyl]amino}-N,N-bis(4-methoxybenzyl)hexanamide;3-{[bis(2-thienylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)propanamide;3-{butyl[(2-thienylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)propanamide;benzyl{(5S)-6-[bis(2-thienylmethyl)amino]-5-cyano-6-oxohexyl}carbamate;benzyl{(5R)-5-azido-6-[bis(2-thienylmethyl)amino]-6-oxohexyl}carbamate;benzyl{(5S)-6-[bis(3-thienylmethyl)amino]-5-[(tert-butoxycarbon-yl)amino]-6-oxohexyl}carbamate;and mixtures or combinations thereof.

In other embodiments, a method of enhancing binding of cells to anintegrin-binding ligand is provided, wherein an agonist of integrin is acompound selected from the group consisting ofN-(3-methoxybenzyl)-N,N′,N′-tris(2-thienylmethyl)pentanediamide;N-[2-(2-thienyl)ethyl]-N,N′,N′-tris(2-thienylmethyl)pentanediamide;N,N-bis(3-methoxybenzyl)-N′,N′-bis (2-thienylmethyl)pentanediamide;N,N-bis(pyridin-4-ylmethyl)-N′,N′-bis (2-thienylmethyl)pentanediamide;N,N,N′,N′-tetrakis(2-thienylmethyl)hexanediamide;N,N,N′,N′-tetrakis(3-methoxybenzyl)hexanediamide;N,N,N′,N′-tetrakis(4-methoxybenzyl) hexanediamide;(3E)-N,N,N′,N′-tetrakis(2-thienylmethyl)hex-3-enediamide;N,N,N′,N′-tetrakis(2-thienylmethyl)pentanediamide;N,N,N′,N′-tetrakis(4-methoxybenzyl) pentanediamide;2,2′-oxybis[N,N-bis(2-thienylmethyl)acetamide]; N,N,N′,N′-tetrakis(2-thienylmethyl)octanediamide;N,N,N′,N′-tetrakis(2-thienylmethyl)heptanediamide;3-oxo-1-(2-thienyl)-2-(2-thienylmethyl)-4,7,10-trioxa-2-azadodecan-12-ylbis(2-thienylmethyl)carbamate;2,2′-(1,3-phenylene)bis[N,N-bis(2-thienylmethyl)acetamide];N,N,N′,N′-tetrakis(4-methoxybenzyl)heptanediamide;N,N,N′,N′-tetrakis(4-methoxybenzyl)succinamideethane-1,2-diylbis[bis(2-thienylmethyl)carbamate];N,N,N′,N′-tetrakis(4-methoxybenzyl)octanediamide;N,N,N′,N′-tetrakis(2-thienylmethyl) pyridine-3,5-dicarboxamide;N,N,N′,N′-tetrakis(2-thienylmethyl)pyridine-2,6-dicarboxamide;N,N,N′,N′-tetrakis(2-thienylmethyl)pyridine-2,4-dicarboxamide;2,2′-(1,4-phenylene)bis [N,N-bis(2-thienylmethyl)acetamide];N,N′-bis(4-methoxybenzyl)-N,N′-bis (2-thienylmethyl)hexanediamide; andmixtures or combinations thereof.

In another embodiment, a method of enhanced binding ofintegrin-expressing cells to an integrin-binding ligand utilizes anintegrin agonist compound selected from the group consisting ofmethyl(2S)-6-{[(benzyloxy)carbonyl]amino}-2-[benzyl(phenylsulfonyl)amino]hexanoate;methyl(2S)-6-{[(benzyloxy)carbonyl]amino}-2-[benzyl(2-thienylsulfonyl)amino]hexanoate;methyl(2S)-6-{[(benzyloxy)carbonyl]amino}-2-[(2-thienylacetyl)(2-thienylmethyl)amino]hexanoate;methyl(2S)-6-{[(benzyloxy)carbonyl]amino}-2-[(2-thienylcarbonyl)(2-thienylmethyl)amino]hexanoate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(2-thienylmethyl)(2-thienylsulfonyl)amino]hexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(phenylsulfonyl)(2-thienylmethyl)amino]hexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(2-thienylacetyl)(2-thienylmethyl)amino]hexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(methylsulfonyl)(2-thienylmethyl)amino]hexyl}carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(2-thienylcarbonyl)(2-thienylmethyl)amino]hexyl}carbamate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-{[(4-methoxyphenyl)sulfonyl]-(2-thienylmethyl)amino}hexyl]carbamate;benzyl{(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(4-methoxybenzoyl)(2-thienylmethyl)amino]hexyl}carbamate;N,N′-heptane-1,7-diylbis[N-(2-thienylmethyl)thiophene-2-carboxamide];N,N′-heptane-1,7-diylbis[N-(2-thienylmethyl)benzamide;N,N′-hexane-1,6-diylbis [N-(2-thienylmethyl)thiophene-2-carboxamide];N,N′-hexane-1,6-diylbis[N-(3-methoxybenzyl) thiophene-2-carboxamide];tert-butyl{5-[(4-methoxybenzyl)(2-thienylsulfonyl)amino]pentyl}carbamate;N-(3-methoxybenzyl)-N-{5-[(2-thienylsulfonyl)amino]pentyl}thiophene-2-sulfonamide;tert-butyl{(2S)-1,6-bis[bis(2-thienylmethyl)amino]-1,6-dioxohexan-2-yl}carbamate;tert-butyl{5-[(2-thienylcarbonyl)(2-thienylmethyl)amino]pentyl}carbamate;N-(3-methoxybenzyl)-N-{5-[(2-thienylcarbonyl)amino]pentyl}thiophene-2-carboxamide;N,N′-pentane-1,5-diylbis[N-(3-methoxybenzyl)thiophene-2-carboxamide];and mixtures or combinations thereof.

In a further embodiment, a method of enhanced binding ofintegrin-expressing cells to an integrin-binding ligand utilizes anintegrin agonist compound selected from the group consisting ofN-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}-N-(2-thienylmethyl)thiophene-2-carboxamide;2-{butyl[(2-thienylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)ethanesulfonamide;2-[(methylsulfonyl)(2-thienylmethyl)amino]-N,N-bis(2-thienylmethyl)ethanesulfonamide;2-{[bis(2-thienylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)ethanesulfonamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}thiophene-2-sulfonamide;N-{2-[bis(2-thienylmethyl) sulfamoyl]ethyl}-2-(2-thienyl)acetamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}thiophene-2-carboxamide;N,N-bis(2-thienylmethyl)-2-{[(2-thienylmethyl)carbamoyl]amino}ethanesulfonamide;2-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)acetamide;3-[{2-[bis(2-thienylmethyl)amino]-2-oxoethyl}(butyl)amino]-N,N-bis(2-thienylmethyl)propanamide;2-[{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}(methyl)amino]-N,N-bis(2-thienylmethyl)acetamide;3-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)propanamide;3-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(4-methoxybenzyl)propanamide;2-(acetyl{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)acetamide;2-(acetyl{2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)acetamide; and mixtures or combinations thereof.

In some embodiments, a method of enhancing binding of cells to anintegrin-binding ligand is provided, wherein said agonist of integrin isa compound selected from the group consisting oftert-butyl[(2S)-1-{[bis(cyclopropylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;(2S)-[(tert-butoxycarbonyl)amino]hexyldiisobutylcarbamate;methyl(8S,12S)-12-(1,3-benzodioxol-5-yl)butyl-4-isobutyl-2-methyl-5,10-dioxo-6-oxa-4,9,11-triazatetradecan-14-oate;benzyl{(5S)-6-[bis(cyclopropylmethyl)amino]-5-[(tert-butoxycarbonyl)amino-]-6-oxohexyl}arbamate;and mixtures or combinations thereof.

Vaccines

Background

During the priming and effector phases of the immune response, antigensare taken up by cells termed antigen presenting cells (APCs). APCs thenpresent this antigen in the context of major histocompatibility complex(MHC) molecules on their cell surface, to stimulate lymphocytes tobecome activated, proliferate, and differentiate into effector cellsthat can eventually protect the host. CD4⁺ T-helper 1 cells (Th1 cells)directly interact with APCs and produce cytokines, which drivelymphocyte proliferation and the eventual development of memory T cells,and effector cytotoxic T Lymphocytes (CTLs). CD4⁺ T-helper 2 cells (Th2cells) directly interact with antigen presenting cells and drive B-cellactivation and proliferation, eventually resulting in the production ofa humoral, antibody-dependent immune response. Communication between avariety of cell types must occur through direct cell contact for aproductive immune response to follow antigenic challenge. Directcell-cell contacts are mediated by the integrin family of cell adhesionmolecules.

Integrin cell adhesion molecules are cell surface glycoproteinscomprised on non-covalently associated α and β heterodimers. Eighteendifferent α-chains combine with 8 β-chains to form 24 different αβpairs. On leukocytes, integrins are intimately involved in the adhesioncascade, which governs leukocyte trafficking to sites of inflammation orinjury. Integrins are also key components in the generation of anadaptive immune response, which is easily observed during the process ofvaccination.

Integrins are also essential in the priming phase of the immuneresponse, as this phase requires the direct interaction betweenlymphocytes and APCs. The integrins α4β1 and αLβ2 both have been shownto be important in lymphocyte conjugation with APCs and have been shownto be able to provide costimulatory signals that result in lymphocyteactivation, proliferation, differentiation and even positive selection.Fibronectin promotes proliferation of naive and memory T cells bysignaling through both the VLA-4 and VLA-5 integrin molecules.Activating these integrins to promote efficient conjugation betweenlymphocytes and APCs would augment the immune response by enhancingintegrin costimulatory effects. For a number of diseases, currentvaccination strategies are inadequate to generate long term protectiveimmunity. This is especially true in the case of vaccines directedtowards tumor-associated antigens in cancer. We have now discovered thatsmall molecule compounds that promote integrin:ligand interactions, suchas those embodied in United States Published Patent Application No.20130236434A1 and Vanderslice, P., Biediger, R. J., Woodside, D. G.,Brown, W. S., Khounlo, S., Warier, N. D., Gundlach, C. W. t., Caivano,A. R., Bornmann, W. G., Maxwell, D. S., et al. “Small molecule agonistof very late antigen-4 (VLA-4) integrin induces progenitor celladhesion,” J Biol Chem 288, 19414-19428 2013(Vanderslice et al. 2013),effectively function as: (1) targeted adjuvant to safely increase thepriming responses with current vaccination approaches, and (2) increaseeffector functions of both humoral and innate immune responses. Biedigeret al and Vanderslice et al failed to recognize the use of such integrinagonists as immunomodulators which could be used to drive many aspectsof the immune response independent of their ability to cause increasedretention within target tissues. In examining these integrin agonistsfurther, we discovered that their use could be expanded to multipleaspects of the immune response, which are described below.

Vaccines Embodiments

A pharmaceutical preparation of a vaccine comprising an antigen andeffective amount of one or a plurality of enhancing compounds, where theassociated enhancing compounds are capable of enhancingintegrin-mediated binding of integrins of a cell to their respectiveligand, wherein the integrins include α4β1, α4β7, α5β1, and/or αLβ2,wherein the ligands include VCAM-1, fibronectin, MAdCAM-1, ICAM-1,and/or ICAM-2, wherein the antigen is a purified protein, peptide, cell,or cell lysate.

A pharmaceutical preparation of an anticancer vaccine comprising anantigen and effective amount of one or a plurality of enhancingcompounds, where the associated enhancing compounds are capable ofenhancing integrin-mediated binding of integrins of a cell to theirrespective ligand, wherein the integrins include α4β1, α4β7, α5β1,and/or αLβ2, wherein the ligands include VCAM-1, fibronectin, MAdCAM-1,ICAM-1, and/or ICAM-2, wherein the antigen is a purified protein,peptide, tumor cell, or tumor lysate.

A pharmaceutical preparation of an anti-cancer vaccine comprising anantigen, adjuvant and effective amount of one or a plurality ofenhancing compounds, where the associated enhancing compounds arecapable of enhancing integrin-mediated binding of integrins of a cell totheir respective ligand, wherein the integrins include α4β1, α4β7, α5β1,and/or αLβ2, wherein the ligands include VCAM-1, fibronectin, MAdCAM-1,ICAM-1, ICAM-2, wherein the antigen is a purified protein, peptide,tumor cell, or tumor lysate, wherein the adjuvant is non-specific orspecific substance capable of eliciting an immune response in responseto an antigen. Examples of non-specific adjuvants include BCG, completefreund's adjuvant, alum, or noscapine. Specific adjuvants includewithout limitations G-CSF, FGF, Toll-like receptor agonists, immunecheckpoint inhibitors (CTL-4, PD-1, PDL-1, IDO-1). The integrinenhancing compounds are considered specific adjuvants.

Method to treat a patient with an effective amount of one or a pluralityof enhancing compounds, wherein the associated enhancing compounds arecapable of enhancing integrin-mediated binding of integrins of a cell totheir respective ligand, wherein the integrins include α4β1, α4β7, α5β1,and/or αLβ2, wherein the ligands include VCAM-1, fibronectin, MAdCAM-1,ICAM-1, and/or ICAM-2, in combination with a therapeutically ineffectivedose of immune checkpoint inhibitor, cytotoxic chemotherapy, cancervaccine, wherein the patient has been diagnosed with cancer.

Method to treat a patient with an effective amount of one or a pluralityof enhancing compounds, wherein the associated enhancing compounds arecapable of enhancing integrin-mediated binding of integrins of a cell totheir respective ligand, wherein the integrins include α4β1, α4β7, α5β1,and/or αLβ2, wherein the ligands include VCAM-1, fibronectin, MAdCAM-1,ICAM-1, and/or ICAM-2, in combination with a therapeutically ineffectivedose of immune checkpoint inhibitor, cytotoxic chemotherapy, cancervaccine, wherein the patient has been diagnosed with solid tumors in thelung, prostate, breast, colon, skin, brain, or pancreas, wherein theenhancing compound is systemic administered by enteral or parenteralroute of administration.

Method to treat a patient with an effective amount of one or a pluralityof enhancing compounds, where the associated enhancing compounds arecapable of enhancing integrin-mediated binding of integrins of a cell totheir respective ligand, wherein the integrins include α4β1, α4β7, α5β1,and/or αLβ2, wherein the ligands include VCAM-1, fibronectin, MAdCAM-1,ICAM-1, ICAM-2, and/or vitronectin, in combination with atherapeutically ineffective dose of immune checkpoint inhibitor,cytotoxic chemotherapy, wherein the patient has been diagnosed withhematologic cancers, wherein the integrin mimetic is systemicadministered by enteral or parenteral route of administration.

Adoptive Cell Therapy

Background

One of the outcomes of a productive immune response is the killing ofspecific target cells by effector lymphocytes. Antigen specific CTLs,Natural Killer (NK) cells, invariant NK T (iNKT) cells, and variousengineered derivatives of these cells, all of which are under clinicalinvestigation as cancer therapies, bind to and directly kill theirintended cellular targets. Conjugate formation between CTLs or NK cellsand their targets requires cell adhesion mediated by the integrin familyof cell adhesion molecules, in particular the integrins α4β1 and αLβ2.Cross-linking of α4β1 and α5β1 fibronectin receptors enhances naturalkiller cell cytotoxic activity. By enhancing these adhesive andcostimulatory interactions, activators of integrin adhesion will promotethis cytolytic activity and increase the in vivo killing in both vaccinestrategies, and in adoptive cell therapies. This is exemplified in FIGS.1&2. The pharmaceutical compositions comprising an enhancing compoundalone in a vaccine based strategy, or in a combination with effectorcells could be particularly useful in patients with refractory cancers.

Adoptive Cell Therapy Embodiments

A pharmaceutical preparation comprising NK cells, activated NK cells,engineered NK cells, or NK cell lines (and derivatives thereof) treatedwith an effective amount of one or a number of compounds capable ofactivating integrin mediated interactions with their cognate ligands.

A method to treat a patient with a pharmaceutical preparation comprisingNK cells, activated NK cells, engineered NK cells, or NK cell lines (andderivatives thereof) delivered in combination with (either prior to,during, or after) an effective amount of one or a number of compoundscapable of activating integrin mediated interactions with their cognateligands.

A method to treat a patient with a pharmaceutical preparation comprisinga therapeutically ineffective amount of NK cells, activated NK cells,engineered NK cells, or NK cell lines (and derivatives thereof)delivered in combination with (either prior to, during, or after) aneffective amount of one or a number of compounds capable of activatingintegrin mediated interactions with their cognate ligands.

A pharmaceutical preparation comprising invariant natural killer T(iNKT) cells, activated iNKT cells, engineered iNKT cells, or iNKT celllines (and derivatives thereof) treated with an effective amount of oneor a number of compounds capable of activating integrin mediatedinteractions with their cognate ligands.

A method to treat a patient with a pharmaceutical preparation comprisingiNKT cells, activated iNKT cells, engineered iNKT cells, or iNKT celllines (and derivatives thereof) delivered in combination with (eitherprior to, during, or after) an effective amount one or a number ofcompounds capable of activating integrin mediated interactions withtheir cognate ligands.

A method to treat a patient with a pharmaceutical preparation comprisinga therapeutically ineffective amount of iNKT cells, activated iNKTcells, engineered iNKT cells, or iNKT cell lines (and derivativesthereof) delivered in combination with (either prior to, during, orafter) an effective amount one or a number of compounds capable ofactivating integrin mediated interactions with their cognate ligands.

A pharmaceutical preparation comprising γδT cells, activated iNKT cells,engineered γδT cells, or γδT cell lines (and derivatives thereof)treated with an effective amount of one or a number of compounds capableof activating integrin mediated interactions with their cognate ligands.

A method to treat a patient with a pharmaceutical preparation comprisingγδT cells, activated iNKT cells, engineered γδT cells, or γδT cell lines(and derivatives thereof) delivered in combination with (either priorto, during, or after) an effective amount one or a number of compoundscapable of activating integrin mediated interactions with their cognateligands.

A method to treat a patient with a pharmaceutical preparation comprisingγδT cells, activated iNKT cells, engineered γδT cells, or gδT cell lines(and derivatives thereof) delivered in combination with (either priorto, during, or after) an effective amount one or a number of compoundscapable of activating integrin mediated interactions with their cognateligands.

A pharmaceutical preparation comprising cytotoxic T lymphocytes (CTLs)(including, but not limited to, tumor infiltrating lymphocytes,lymphocytes expanded in an antigen specific manner, lymphocytesengineered to express chimeric antigen receptors) treated with aneffective amount of one or a number of compounds capable of activatingintegrin mediated interactions with their cognate ligands for thepurpose of enhancing CTL tumoricidal activity.

A method to treat a patient with a pharmaceutical preparation comprisingcytotoxic T lymphocytes (CTLs) (including, but not limited to, tumorinfiltrating lymphocytes, lymphocytes expanded in an antigen specificmanner, lymphocytes engineered to express chimeric antigen receptors)delivered in combination with (either prior to, during, or after) aneffective amount of one or a number of compounds capable of activatingintegrin mediated interactions with their cognate ligands for thepurpose of enhancing CTL tumoricidal activity.

A method to treat a patient with a pharmaceutical preparation comprisinga therapeutically ineffective amount of cytotoxic T lymphocytes (CTLs)(including, but not limited to, tumor infiltrating lymphocytes,lymphocytes expanded in an antigen specific manner, lymphocytesengineered to express chimeric antigen receptors) delivered incombination with (either prior to, during, or after) an effective amountof one or a number of compounds capable of activating integrin mediatedinteractions with their cognate ligands for the purpose of enhancing CTLtumoricidal activity.

Antibody Dependent Cellular Cytotoxicity

Background

Antibody dependent cellular cytotoxicity (ADCC) is a mechanism wherebyantibodies bind a cell surface antigen, opsonizing the cell forengulfment (phagocytosis) and/or destruction by effector cells of theimmune system such as neutrophils, macrophage, and NK cells that expressactivating receptors for the Fc components of antibodies. The in vivomechanism of action of humanized monoclonal antibodies (mAbs)trastuzumab (Herceptin®), rituximab (Rituxan®), ipilimumab (Yervoy@) andothers involve ADCC. The β2 integrin family (including integrinCD11a/CD18 (also called αLβ2 or LFA-1) are important mediators of thedirect binding between effectors and antibody-opsonized target cellsduring ADCC. Enhancing ADCC with small molecule integrin activatorswould promote ADCC and serve as an adjunct therapy with any therapeuticantibody with an Fc component.

Pharmaceutical compositions comprising an enhancing compound incombination with a therapeutic antibody is useful in patients to treatminimal residual disease, and those refractory to such therapeuticantibodies.

ADCC Embodiments

A method to treat a patient with a therapeutic antibody, wherein theantibody contains a Fc region, wherein the antibody is, withoutlimitation, trastuzumab, cetuximab, ipilimumab, nivolumab, rituximab,alemtuzumab, atumumab, or tositumomab, and an effective amount of one ora number of compounds capable of activating integrin mediatedinteractions with their cognate ligands.

A method to treat a patient with therapeutically ineffective amount oftherapeutic antibody, wherein the antibody contains a Fc region, whereinthe antibody is, without limitation, trastuzumab, cetuximab, ipilimumab,nivolumab rituximab, alemtuzumab, atumumab, or tositumomab, and aneffective amount of one or a number of compounds capable of activatingintegrin mediated interactions with their cognate ligands.

A pharmaceutical composition comprising a therapeutically ineffectiveamount of therapeutic antibody, wherein the antibody contains a Fcfragment and an effective amount of one or a number of compounds capableof activating integrin mediated interactions with their cognate ligands.

TABLE I Compound Designation, Names, and Structures Compound C1(6S,10S)-methyl 10-1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate C2 N¹,N¹,N⁶,N⁶-tetrakis(4-methoxybenzyl)adipamideC3 N¹,N¹,N⁶,N⁶-tetrakis(3-methoxybenzyl)adipamide C4(ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl)bis(bis(thiophen-2-ylmethyl)carbamate)

C3

Antibody dependent cellular cytotoxicity (ADCC) is mediated by naturalkiller (NK) cells, monocytes and macrophage, and other effector cellsthat express receptors (like CD16) that can bind to antibodies bound totarget cells.

Cell adhesion between effector cells like NK cells and their targetcells is mediated by integrin cell adhesion molecules. This adhesion isessential for both cytolytic killing activity and ADCC. Compounds suchas C4 and other compounds of Formula (I), target integrins α4β1 andαLβ2, which are essential in cytotolytic T cell and NK cell dependentkilling of tumor targets, and are required for ADCC in vivo.

C4 and/or other compounds of Formula (I) are positive allostericmodulators of integrin cell adhesion molecules, inducing integrins α4β1and αLβ2 (among others), to interact with their cognate ligands (TableII).

TABLE II Activity (EC₅₀, μM) of C4 in Enhancing Adhesion Mediated byDifferent Classes of Integrins Compound α4β1 (VLA-4) αLβ2 (LFA-1) C1 3226 C2 11 11 C3 11 7 C4 14 11

As such, the compounds can also facilitate chemotactic cell migration onthe α4β1 integrin substrate vascular cell adhesion molecule-1 (VCAM-1)and the αLβ2 integrin substrate intercellular adhesion molecule-1(ICAM-1) as shown in FIGS. 3A&B.

Because C4 can activate cell adhesion, it is predicted to enhance theimmune response through a number of different mechanisms. This could beby augmenting antigen presentation between lymphocytes and antigenpresenting cells. This could also be through increased effector celltrafficking to sites of an immune response. Importantly, augmentedimmune responses could be the result of more efficient cytolyticeffector cell killing of tumor target cells. This could be in thecontext of simple NK cell or CTL mediated killing, or in the context oftarget cell killing via ADCC.

Augmentation of NK Cytolytic Activity by C4

Integrins, including C4 targets α4β1 and αLβ2, are essential in CTL andNK cell dependent killing of tumor targets. Thus, by facilitating celladhesion, C4 should increase tumor target killing by effector cells. Totest this general concept, freshly isolated NK cells, or the NK cellline NK92 were mixed with K562 tumor targets in standard cytotoxicityassays with, or without, C4. C4 enhanced NK and NK92-mediated killing ofK562 tumor target cells see FIGS. 1 and 2. Although the target cells inthis example are erythroleukemic cells, these results are extendible tosolid tumors as well as many of these tumor types express the ligandsfor integrins α4β1 and αLβ2.

Checkpoint Blockade is Mediated by ADCC

Effectiveness of anti-CTLA4 therapy in melanoma patients is due in partto ADCC dependent selective depletion of negative regulatory T cells ina solid tumor. This has also been shown to the case in animal models ofmelanoma. When anti-CTLA-4 mAb (clone 9H10) is administered incombination with a vaccine termed GVAX®, melanoma regression occurs asregulatory T cells are depleted by Fc receptor bearing cells in thetumor microenvironment. As C4 can enhance the cytolytic mechanismsinvolved in ADCC, it was hypothesized that C4 would be able to augmentthe therapeutic effects of anti-CTLA-4 treatment (in particular with theanti-CTLA-4 mAb clone 9H10) in an experimental model of melanoma.

C4 Enhancement of GVAX®/Anti-CTLA-4 Treatment

C4 was tested in a β16-BL6 melanoma model to determine potential effectson the current checkpoint blockade therapy anti-CTLA-4. Briefly, thirtyC57BL/6 were randomized into 3 groups (n=10). C4 was added to a regimenof GVAX®/anti-CTLA-4. Historically, these treatment regimens result inat best, a 20% tumor free survival. On day 0, β16-BL6 cells (2.5×10⁴)were injected subcutaneously. GVAX®/anti-CTLA-4 (clone 9H10), or IgGcontrols, were administered on days 3, 6, and 9. On day 11, C4 treatmentwas initiated intratumorally twice weekly (1 mg/kg) for 30 days. Vehiclecontrol was added to the IgG group. Addition of C4 to theGVAX®/anti-CTLA-4 treatment group demonstrated a significant mediansurvival benefit (Table III) along with a significant increase in longterm tumor free survival (Table IV).

TABLE III C4 Increases Median Survival of Mice with Metastatic Melanomain Combination with Anti-CTLA-4 Median Survival Hazard 95% CI ofTreatment Group (days) Comparisons Ratio Ratio P value Untreated 29.00anti-CTLA-4 32.00 Untreated vs. Anti- 0.32 0.06 to 0.36 <0.05 CTLA-4anti-CTLA-4 + C4 71.50 Anti-CTLA-4 vs. Anti- 0.37 0.10 to 0.90 <0.05CTLA-4 + THI349 Notes: At day 0, C57BL/6 mice were implanted with 2.5 ×10⁴ B16BL6 melanoma cells, and on day 3, 6, 9, treated with Anti-CTLA-4,clone 9H10, Bioxcell, 10 mg/kg) with and without C4 (intratumorally, 1mg/kg, twice weekly for 30 days) in a GVAX ® background. P value byLog-rank, time to event based on tumor burden of 200 mm². The censor dayon the survival data was 100 days.

TABLE IV C4 Increases the Incidence of Long-term Tumor Rejection inCombination with Anti-CTLA-4 in Mice with Metastatic Melanoma # to Pvalue Treatment Tumor free (Chi- Group animals Comparisons square)Untreated  0.0% anti-CTLA-4 11.1% Untreated vs. Anti- 0.14 CTLA-4anti-CTLA-4 + 50.0% Anti-CTLA-4 vs. Anti- 0.03 C4 CTLA-4 + C4 Notes: Pvalue by Chi-square one tailed. Animal were palpated at the tumorinjection site over a 100 day period for the presence of a tumor.

C4 has demonstrated significant increases in median survival times andlong term tumor free survival in combination with the checkpointblockade treatment anti-CTLA-4. Notably, the animal model data presentedabove utilized the same anti-CTLA-4 clone, mAb clone 9H10, which hasbeen shown to produce therapeutic effects in this model by ADCC.

C4 and related compounds of Formula (I), enhance cell adhesion mediatedby integrins that are essential for ADCC. In an in vivo tumor model, C4significantly enhanced the therapeutic efficacy of a CTLA-4 antibody,mAb clone 9H10, which is known to be effective through ADCC. Byextension, C4 and related compounds of Formula (I) are thought to beeffective via enhancement of ADCC.

Antibody dependent cellular cytotoxicity (ADCC) is a mechanism wherebyantibodies bind a cell surface antigen, opsonizing the cell forengulfment (phagocytosis) and/or destruction by effector cells of theimmune system such as neutrophils, macrophage, and NK cells that expressactivating receptors (e.g., Fc receptors) for the Fc components ofantibodies see FIGS. 4A-C.

Referring to FIGS. 4A-C, a mechanism of ADCC is shown. Looking at FIG.4A, a tumor target cell unbound with therapeutic antibody, no ADCCoccurs. Looking at FIG. 4B, an Fc receptor binds to therapeutic antibodybound tumor target cell, but weak adhesion between effector and targetcell results in weak ADCC. Looking at FIG. 4C, the Fc receptor oneffector cell binds to therapeutic antibody bound tumor target, andstrong adhesion mediated by integrins between effector and target cellresults in strong ADCC.

Enhancing ADCC with small molecule integrin activators would promoteADCC and serve as an adjunct therapy with any therapeutic antibody withan Fc component that may mediate ADCC. The results presented heredemonstrate the ability of C4 to augment the therapeutic activity of mAb9H10, which acts in vivo through a mechanism of ADCC. By extension, thismechanism of action could be applied to enhance therapeutic efficacy ofany mAb treatment for which ADCC may occur.

Experiments Relating to ADCC

Reagents and Cell Lines

For all assays described, C4 was dissolved in DMSO to make a 1 mM stocksolution, and dilutions were made in assay buffer or media to yield thedesired final working concentrations in 1% DMSO (vehicle). Human VCAM-1,MAdCAM-1 Fc chimera, ICAM-1, and SDF-la were purchased from R&D Systems(Minneapolis, Minn.). Human serum fibronectin was purchased fromSigma-Aldrich (St. Louis, Mo.). The cell lines NK-92, Jurkat, K562, andHSB were obtained from American Type Culture Collection (Manassas, Va.)and were maintained in recommended culture media. The mutant Jurkat cellline not expressing α4 integrin (Jurkat [α4−]) was a gift from Dr. DavidRose, University of California San Diego, La Jolla, Calif.

Static Cell Adhesion Assays

Ligands (VCAM-1, MAdCAM-1, fibronectin, or ICAM-1) in 50 μL of 50 mMTris-HCl (pH 7.4), 150 mM NaCl, (TBS) were added to wells of a 96-wellplate and allowed to coat overnight at 4° C. In order to maximize thewindow to evaluate agonist activity, a sub-optimal coating concentrationof ligand was used. This ligand concentration corresponded approximatelyto that which would yield ≤5% adhesion as determined by dose-responsecurves of ligand binding to the appropriate cell type. All assays wereperformed as previously described. Briefly, 2×10⁶ cells were labeled for30 minutes with calcein-AM (Molecular Probes), washed, resuspended inbinding buffer, and added to ligand-coated plates (2×10⁵ cells/well)that had been blocked with 2% BSA. The binding buffer was PBS with 1 mMMgCl₂ and 50% FBS (VCAM-1 and ICAM-1 assays) or TBS with 1 mM MnCl₂ and50% FBS (MAdCAM-1 and fibronectin assays). After a 30-minute incubationat 37° C., the plates were washed 3 times with the respective bindingbuffer (except without the serum), the adherent cells were lysed, andfluorescence was measured on a Tecan Safire²plate reader. Standardcurves were run for each assay to convert fluorescence units to cellnumber. For each assay, the cells expressed the appropriate integrinreceptor either endogenously (Jurkat/α4β1, K562/α5β1, Jurkat (α4−)/αLβ2,HSB/αLβ2) or in recombinant form (K562/437). Generation of therecombinant K562 cell line has been described.

Migration Assays

Migration assays were performed in 3 μM pore size Transwells (24 well,Costar, Cambridge, Mass.). The upper chambers were pre-coated with 10mg/mL VCAM-1 or 1 μg/mL ICAM-1 in 50 mL TBS overnight at 4° C. and werethen blocked with 2% BSA for 1 hour at room temperature. After washingwith migration medium (RPMI-1640 supplemented with 1% FBS, 100 units/mLpenicillin and 100 μg/mL streptomycin), upper chambers were loaded with2×10⁵ cells Jurkat cells (VCAM-1 assays) or Jurkat (α4-) cells (ICAM-1assays) in 160 μL of migration medium. Lower chambers contained 600 μLof migration medium supplemented with 5 μg/mL (VCAM-1 assays) or 1 μg/mL(ICAM-1 assays) SDF-1α to induce chemotaxis. Jurkat cells were mixedwith vehicle (1% DMSO) or C4 at the indicated concentrations immediatelyprior to being added to the upper chamber. After a 4-hour incubation at37° C., 5% CO₂, the upper chambers were removed, and cells in the lowerchamber were collected and counted on a hemocytometer. Results areexpressed as the total number of cells migrated.

Checkpoint Blockade Melanoma Model

The B16-BL6 therapeutic melanoma model is well described. Briefly,B16-BL6 cells (>90% viability) were be resuspended at 2.5×105 cells/mlin HBSS (4° C.). At day 0 (d0), C57BL/6 mice (4-6 wk old, female) wereinoculated (subcutaneous, flank) with 2.5×10⁴ cells (100 μL). At d3,appropriate groups received β16-GM-CSF vaccine (GVAX®). GVAX®preparation followed established procedures. Log-phase growth B16-GM-CSFcells were harvested as described above for β16-BL6 cells, butresuspended at 1×10⁷ cells in HBSS (4° C.). Cells were irradiated (5000rad) and injected into contralateral flanks (100 μL, 1×10⁶ cells). Alsoat d3, all animals received anti-CTLA-4 (mAb 9H10, BioXcell, 200μg/animal, twice weekly). C4 (1 mg/kg in 25 μL 2× weekly intratumoral),or control vehicle (Veh, 25 μL), was administered at dl 1 in appropriategroups. At d12 and every 2 days thereafter, all animals were observedfor tumor growth. Calipers were used to measure perpendicular tumordiameters. Animals were monitored daily for distress and survival wasassessed twice daily. Lack of survival was defined as death, or tumorsize >200 mm².

NK Cytotoxicity Testing

Human NK cells were purified from leukoreduction filters obtained fromhealthy volunteers using a CD56+ magnetic bead based enrichment columnfrom Multenyi Biotech (as per manufacturer's instructions). Purifiedcells were routinely >90% CD56+ positive as measured by flow cytometry.For cytotoxicity testing, logarithmically growing K562 target cells werelabeled with BATDA (bis (acetoxymethyl)2,2′:6′,2″-terpyridine-6,6″-dicarboxylate, Perkin Elmer DELFIA EuTDACytotoxicity Reagent, Catalogue # ADO 116, as per manufacturer'sinstructions). In round bottom 96-well plates, 1×10⁴ K562 tumor targetcells were added in 100 μL of culture media. This was followed by theaddition of NK cells, at indicated effector to target ratios in a volumeof 100 μL of culture media. C4 was then added to wells, at indicatedconcentrations. After a 4 h incubation at 37° C., 5% CO₂, culturesupernatants were removed and liberated TDA(2,2′:6′,2″-terpyridine-6,6″-dicarboxylic acid) was detected by additionof DELFIA solution to form EuTDA, and fluorescence was measured on aTecan Safire 2 multimodal plate reader. For determination of maximalcell lysis, 1.0% TritonX-100 was added to control wells prior to removalof supernatant. For spontaneous release of TDA, labelled K562 wereincubated identically as described above, in the presence of C4, butwithout NK effector cells. Results are expressed as the percent specificlysis, calculated as [(experimental signal−spontaneous release)/(totalsignal−spontaneous release).

All references cited herein are incorporated by reference. Although theinvention has been disclosed with reference to its preferredembodiments, from reading this description those of skill in the art mayappreciate changes and modification that may be made which do not departfrom the scope and spirit of the invention as described above andclaimed hereafter.

We claim:
 1. A method comprising: administering to a subject in needthereof: (A) an antigen, and (B) an adjuvanting amount of at least oneintegrin activating compound that increases an immune response to theantigen.
 2. The method of claim 1, wherein the administering stepinclude: administering the antigen with the at least one integrinactivating compound.
 3. The method of claim 1, wherein, in theadministering step, the integrin is selected from the group consistingof α4β1, α4β7, α5β1, and αLβ2.
 4. The method of claim 1, wherein, in theadministering step, the integrin is selected from the group consistingof α4β1 and αLβ2.
 5. The method of claim 1, wherein, in theadministering step, the at least one integrin activating compoundcomprises one or more compounds of the general Formula (I):R¹-M¹-N(R²)-M²-M³-M⁴-M⁵-M⁶-R³  (I) wherein a first class of thecompounds of Formula (I) is defined by: R¹ is selected from the groupconsisting of aryl and aralkyl, R² is alkyl, aryl, or aralkyl, M¹ isCH₂, M² is CO, M³ is O, S, or NR⁶, R⁶ when present is hydrogen or loweralkyl, M⁴ is absent or CH₂, M⁵ is (CR¹¹R¹²), R¹¹ is hydrogen, R¹² isselected from the group consisting of hydrogen, NR²¹CONR²²R²³,NR²¹COR²⁴, NR²¹SO₂R²⁴, NR²¹COOR²⁴, OCOR²⁴, OR²⁴, O(CH₂CH₂O)_(s)R²⁴,COOR²⁴, alkyl, and hydroxyalkyl, s is an integer of 1 to 6, R²¹ and R²²when present are independently selected from the group consisting ofhydrogen or lower alkyl, R²³ when present is selected from the groupconsisting of hydroxyalkyl, alkoxyalkyl, alkyl, aryl, aralkyl andalkoxycarbonylalkyl, provided that when M³ is NR⁶ and M⁴ is absent, thenR²³ is not 1-(1,3-benzodioxol-5-yl)-3-ethoxy-3-oxopropyl, R²⁴ whenpresent is selected from the group consisting of alkyl, aryl, aralkyl,heterocyclyl, cycloalkyl, cycloalkylalkyl, and heterocyclylalkyl, andmixtures thereof, M⁶ is (CH₂)_(q), where q is an integer from 0 to 6, R³is selected from the group consisting of hydrogen, CONR¹³R¹⁴,NR¹⁵COOR¹⁶, NR¹⁵COR¹⁶, NR¹⁵CONR¹³R¹⁴, NR¹⁵SO₂R¹⁶, OCOR¹⁶, COOR¹⁶, OR¹⁶,SR¹⁶, heterocyclyl, hydroxyl, hydroxyalkyl, guanadino, alkyl and aryl,R¹³ and R¹⁵ when present are independently hydrogen or lower alkyl, R¹⁴and R¹⁶ when present are independently selected from the groupconsisting of hydrogen, alkyl, aryl, aralkyl, cycloalkyl, heterocyclyl,cycloalkylalkyl, and heterocyclylalkyl, R¹, R², R³, R¹², R¹⁴, R¹⁶, R²³and R²⁴ when present may independently be either unsubstituted orsubstituted with one or more substituents selected from the groupconsisting of alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl,heterocyclyl, heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy,azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl, halo,haloalkyl, haloaryl, amino, alkylamino, dialkylamino, arylamino,diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl),—NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl), —NHSO₂(aralkyl),alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino),—OCO(dialkylamino), and mixtures thereof; or wherein a second class ofthe compounds of Formula (I) is defined by: R¹ is aryl or aralkyl, R² isalkyl or aralkyl, M¹ is CH₂, M² is CO, M³ is absent or is O or CH₂, M⁴is absent or is CH₂, M⁵ is absent or is O or (CR¹¹R¹²), R¹¹ is hydrogen,R¹² is selected from the group consisting of hydrogen, NR²¹CONR²²R²³,NR²¹COR²⁴, NR²¹SO₂R²⁴ and NR²¹COOR²⁴, R²¹ and R²² each of which, whenpresent is independently selected from the group of hydrogen and loweralkyl, R²³ and R²⁴, each of which, when present is independentlyselected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, M⁶ isselected from the group consisting of (CH₂)_(q),(CH₂)_(q)—CH═CH—(CH₂)_(r), (CH₂)_(q)-arylene-(CH₂)_(r) and(CH₂CH₂O)_(q), where q and r are independently integers from 0 to 6, R³is CONR¹³R¹⁴, R¹³ and R¹⁴, each of which, when present is independentlyselected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, and R¹,R², R¹³, R¹⁴, R²³ and R²⁴, when present, independently either areunsubstituted or are substituted with one or more substituents selectedfrom the group consisting of alkyl, aryl, aralkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl,hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl,alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino,arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl),—NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl), —NHSO₂(aralkyl),alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino),—OCO(dialkylamino), and mixtures thereof; or wherein a third class ofthe compounds of Formula (I) is defined by: R¹ is aryl or aralkyl, R² isalkyl or aralkyl, M¹ is CH₂, M² is SO₂ or CO, M³ is absent or is CH₂, M⁴is absent or is CH₂, M⁵ is absent or is (CR¹¹R¹²), R¹¹, when present, ishydrogen, R¹², when present, is selected from the group consisting ofhydrogen, alkyl, NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴ and NR²¹COOR²⁴,R²¹ and R²², each of which when present, is independently selected fromthe group of hydrogen, lower alkyl, and aralkyl, R²³ and R²⁴, each ofwhich, when present is independently selected from the group consistingof hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl,heterocyclylalkyl and aralkyl, M⁶ is (CH₂)_(q), or NR³⁴(CH₂)_(q), q isan integer from 0 to 6, R³⁴, when present, is selected form the groupconsisting of alkyl, aralkyl, COR³⁵, and SO₂R³⁵, R³⁵ when present, isselected form the group consisting of alkyl, aryl, and aralkyl, and R³is selected from the group consisting of CONR¹³R¹⁴, SO₂NR¹³R¹⁴,NR¹⁵COOR¹⁶, NR¹⁵COR¹⁶, NR¹⁵CONR¹³R¹⁴, and NR¹⁵SO₂R¹⁶, R¹³ and R¹⁴, eachof which, when present, is independently selected from the groupconsisting of hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl,heterocyclyl, heterocyclylalkyl and aralkyl, R¹⁵ and R¹⁶, each of whichwhen present, is independently selected from the group of hydrogen,lower alkyl, and aralkyl, R¹, R², R¹³, R¹⁴, R¹⁵, R¹⁶, R²³, R²⁴, R³⁴ andR³⁵, when present, either are unsubstituted or are substituted with oneor more substituents selected from the group consisting of alkyl, aryl,aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl,heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl,aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino,alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl),—NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO₂(alkyl),—NHSO₂(aryl), —NHSO₂(aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl,—OCO(alkylamino), and —OCO(dialkylamino), with the proviso that when M²is CO, then M⁶ is NR³⁴(CH₂)_(q), q is not 0; or wherein a four class ofthe compounds of Formula (I) is defined by: R¹ is alkyl, aryl oraralkyl, R² is selected from the group consisting of aralkyl and alkyl,provided that when R¹ is alkyl, R² is aralkyl, M¹ is CO or SO₂, providedthat when M¹ is SO₂ and R¹ is phenyl, 4-methylphenyl or2,4,6-trimethylphenyl, R² is not alkyl, 2-phenethyl, benzyl, or2-methoxy-2-oxoethyl, and when M¹ is CO and R¹ is 2-furyl, 4-pyridyl, or3,5-dinitrophenyl, R² is not alkyl, benzyl or 2-(1H-indol-2-yl)ethyl, M²is absent or CH₂, M³ and M⁴ are absent, M⁵ is (CR¹¹R¹²), R¹¹ ishydrogen, R¹² is selected from the group consisting of hydrogen,NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴, NR²¹COOR²⁴, CONR²²R²³, COOR²⁴,O(CH₂CH₂O)_(s)R²⁴, hydroxyalkyl, and alkoxyalkyl, R²¹, and R²², whenpresent, are independently selected from the group consisting ofhydrogen and C₁-C₆ alkyl, and R²³ and R²⁴, each of which, when present,is independently selected from the group consisting of hydrogen, alkyl,aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl andaralkyl, and s is an integer of 1 to 6, M⁶ is (CH₂)_(q), q is an integerof 0 to 6, R³ is selected from the group consisting of NR¹⁵COOR¹⁶,NR¹⁵COR¹⁶, NR¹⁵CONR¹³R¹⁴, and NR¹⁵SO₂R¹⁶, and R¹³ when present, isindependently selected from the group consisting of hydrogen and C₁-C₆alkyl, and R¹⁴, R¹⁵, and R¹⁶ each of which, when present, areindependently selected from the group consisting of hydrogen, alkyl,aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl andaralkyl, and R¹, R², R³, R¹², R¹⁴, R¹⁵, R¹⁶, R²³, and R²⁴, when present,independently either are unsubstituted or are substituted with one ormore substituents selected from the group consisting of alkyl, aryl,aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl,heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl,aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino,alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl),—NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO₂(alkyl),—NHSO₂(aryl), —NHSO₂(aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl,—OCO(alkylamino) and —OCO(dialkylamino), or pharmaceutically acceptablesalts thereof, or mixtures thereof.
 6. The method of claim 5, wherein,in the administering step, R¹M¹, R², R¹³, and R¹⁴ are independentlyselected from the group consisting of 2-thienylmethyl, 3-methoxybenzyl,4-methoxybenzyl, 2-(2-thienyl)ethyl, pyridin-4-ylmethyl, andpyridin-3-ylmethyl.
 7. The method of claim 6, wherein, in theadministering step: the first class of the compounds are selected fromthe group consisting ofN,N,N′,N′-tetrakis(2-thienylmethyl)pentanediamide;N-(3-methoxybenzyl)-N,N′,N′-tris(2-thienylmethyl)pentanediamide;N,N,N′-tris(2-thienylmethyl)pentanediamide;N′-[2-(2-thienyl)ethyl]-N,N-bis(2-thienylmethyl)pentanediamide;N-[2-(2-thienyl)ethyl]-N,N′,N′-tris(2-thienylmethyl)pentanediamide;N,N-bis(pyridin-4-ylmethyl)-N′,N′-bis(2-thienylmethyl)pentanediamide;N,N-bis(pyridin-3-ylmethyl)-N′,N′-bis(2-thienylmethyl)pentanediamide;N,N-bis(3-methoxybenzyl)-N′,N′-bis(2-thienylmethyl)pentanediamide;N,N,N′,N′-tetrakis(4-methoxybenzyl)pentanediamide;N,N,N′,N′-tetrakis(2-thienylmethyl)hexanediamide;N,N,N′,N′-tetrakis(4-methoxybenzyl) hexanediamide;N,N,N′,N′-tetrakis(3-methoxybenzyl)hexanediamide; N,N,N′,N′-tetrakis(2-thienylmethyl) heptanediamide;2,2′-(1,3-phenylene)bis[N,N-bis(2-thienylmethyl)acetamide];N,N,N′,N′-tetrakis(4-methoxybenzyl)heptanediamide; N,N,N′,N′-tetrakis(2-thienylmethyl)octanediamide; (3E)-N,N,N′,N′-tetrakis(2-thienylmethyl)hex-3-enediamide;2,2′-oxybis[N,N-bis(2-thienylmethyl)acetamide];3-oxo-1-(2-thienyl)-2-(2-thienylmethyl)-4,7,10-trioxa-2-azadodecan-12-ylbis(2-thienylmethyl)carbamate; N,N,N′,N′-tetrakis(4-methoxybenzyl)succinamideethane-1,2-diyl bis[bis(2-thienylmethyl)carbamate];N,N,N′,N′-tetrakis (4-methoxybenzyl)octanediamide;N,N,N′,N′-tetrakis(2-thienylmethyl) pyridine-3,5-dicarboxamide;N,N,N′,N′-tetrakis (2-thienylmethyl)pyridine-2,6-dicarboxamide;N,N,N′,N′-tetrakis (2-thienylmethyl)pyridine-2,4-dicarboxamide;2,2′-(1,4-phenylene) bis[N,N-bis(2-thienylmethyl)acetamide]; 8-{2-[bis(2-thienylmethyl)amino]-2-oxoethoxy}-N,N-bis(2-thienylmethyl)quinoline-2-carboxamide;N,N′-bis(4-methoxybenzyl)-N,N′-bis(2-thienylmethyl)hexanediamide; andtert-butyl{(2S)-1,6-bis[bis(2-thienylmethyl)amino]-1,6-dioxohexan-2-yl}carbamate;the second class of the compounds are selected from the group consistingof 1,2-bis(bis(thiophen-2-ylmethyl)carbamate)ethane;1,2-bis(bis(3-methyloxybenzyl)carbamate)ethane;1,2-bis(bis(4-methyloxybenzyl)carbamate)ethane;1,2-bis((thiophen-2-ylmethyl)(3-methyloxybenzyl)carbamate)ethane;1,2-bis((thiophen-2-ylmethyl)(4-methyloxybenzyl)carbamate)ethane;1,2-bis((3-methyloxybenzyl)(4-methyloxybenzyl)carbamate)ethane;1,5-bis(bis(thiophen-2-ylmethyl)carbamate)-3-oxapentane;1,5-bis(bis(3-methyloxybenzyl)carbamate)-3-oxapentane;1,5-bis(bis(4-methyloxybenzyl)carbamate)-3-oxapentane;1,5-bis((thiophen-2-ylmethyl)(3-methyloxybenzyl)carbamate)-3-oxapentane;1,5-bis((thiophen-2-ylmethyl)(4-methyloxybenzyl)carbamate)-3-oxapentane;1,5-bis((3-methyloxybenzyl)(4-methyloxybenzyl)carbamate)-3-oxapentane;(ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl)bis(bis(thiophen-2-ylmethyl)carbamate)or 1,8-bis(bis(thiophen-2-ylmethyl)carbamate)-3,6-dioxaoctane;1,8-bis(bis(3-methyloxybenzyl)carbamate)-3,6-dioxaoctane;1,8-bis(bis(4-methyloxybenzyl)carbamate)-3,6-dioxaoctane;1,8-bis((thiophen-2-ylmethyl)(3-methyloxybenzyl)carbamate)-3,6-dioxaoctane;1,8-bis((thiophen-2-ylmethyl)(4-methyloxybenzyl)carbamate)-3,6-dioxaoctane;1,8-bis((3-methyloxybenzyl)(4-methyloxybenzyl)carbamate)-3,6-dioxaoctane;1,11-bis(bis(thiophen-2-ylmethyl)carbamate)-3,6,9-trioxaundecane;1,11-bis(bis(3-methyloxybenzyl)carbamate)-3,6,9-trioxaundecane;1,11-bis(bis(4-methyloxybenzyl)carbamate)-3,6,9-trioxaundecane;1,11-bis((thiophen-2-ylmethyl)(3-methyloxybenzyl)carbamate)-3,6,9-trioxaundecane;1,11-bis((thiophen-2-ylmethyl)(4-methyloxybenzyl)carbamate)-3,6,9-trioxaundecane;1,11-bis((3-methyloxybenzyl)(4-methyloxybenzyl)carbamate)-3,6,9-trioxaundecane;1,14-bis(bis(thiophen-2-ylmethyl)carbamate)-3,6,9,12-tetraoxatetradecane;1,14-bis(bis(3-methyloxybenzyl)carbamate)-3,6,9,12-tetraoxatetradecane;1,14-bis(bis(4-methyloxybenzyl)carbamate)-3,6,9,12-tetraoxatetradecane;1,14-bis((thiophen-2-ylmethyl)(3-methyloxybenzyl)carbamate)-3,6,9,12-tetraoxatetradecane;1,14-bis((thiophen-2-ylmethyl)(4-methyloxybenzyl)carbamate)-3,6,9,12-tetraoxatetradecane;1,14-bis((3-methyloxybenzyl)(4-methyloxybenzyl)carbamate)-3,6,9,12-tetraoxatetradecane;1,17-bis(bis(thiophen-2-ylmethyl)carbamate)-3,6,9,12,15-pentaoxatetradecane;1,17-bis(bis(3-methyloxybenzyl)carbamate)-3,6,9,12,15-pentaoxatetradecane;1,17-bis(bis(4-methyloxybenzyl)carbamate)-3,6,9,12,15-pentaoxatetradecane;1,17-bis((thiophen-2-ylmethyl)(3-methyloxybenzyl)carbamate)-3,6,9,12,15-pentaoxatetradecane;1,17-bis((thiophen-2-ylmethyl)(4-methyloxybenzyl)carbamate)-3,6,9,12,15-pentaoxatetradecane;and1,17-bis((3-methyloxybenzyl)(4-methyloxybenzyl)carbamate)-3,6,9,12,15-pentaoxatetradecane;the third class of the compounds are selected from the group consistingof methyl(6S,10S)-1-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10R)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-7-methyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-9-methyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;ethyl(6S,10R)-1-(1,3-benzodioxol-5-yl)-6-butyl-7-methyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(10S)-10-(1,3-benzodioxol-5-yl)-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-2-methyl-3,8-dioxo-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-{[(1,3-benzodioxol-5-ylmethyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate; methyl(6S,10S)-6-butyl-3,8-dioxo-10-phenyl-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-({[(1S)-1-(1,3-benzodioxol-5-yl)-3-hydroxypropyl]carbamoyl}amino)hexylbis(2-thienylmethyl)carbamate;(2S)-2-[(benzylcarbamoyl)amino]hexylbis(2-thienylmethyl)carbamate;(2S)-2-[(morpholin-4-ylcarbonyl)amino]hexylbis(2-thienylmethyl)carbamate;(2S)-2-{[(3-methoxypropyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate;(2S)-2-{[(2-methoxyethyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate;tert-butyl[(2S)-1-{[bis(2-thienylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;(2S)-2-[(tert-butylcarbamoyl)amino]hexyl bis(2-thienylmethyl)carbamate;(2S)-2-[(isopropylcarbamoyl)amino]hexylbis(2-thienylmethyl)carbamate;(2S)-2-[(methylcarbamoyl)amino]hexyl bis(2-thienylmethyl)carbamate;tert-butyl[(2R)-1-{[bis(2-thienylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;benzyl{(5S)-6-{[bis(2-thienylmethyl)carbamoyl]oxy}-5-[(tert-butoxycarbonyl)amino]hexyl}carbamate;methyl(9S,13S)-13-(1,3-benzodioxol-5-yl)-9-({[bis(2-thienylmethyl)carbamoyl]oxy}methyl)-3,11-dioxo-1-phenyl-2-oxa-4,10,12-triazapentadecan-15-oate;(2S)-2-acetamidohexylbis(2-thienylmethyl)carbamate;methyl(6R,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6R,10R)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl{[bis(2-thienylmethyl)carbamoyl](methyl)amino}acetate;methyl{[bis(2-thienylmethyl)carbamoyl](butyl)amino}acetate;tert-butyl{[bis(2-thienylmethyl)carbamoyl](butyl)amino}acetate;benzyl{(5S)-6-{[bis(4-methoxybenzyl)carbamoyl]oxy}-5-[(tert-butoxycarbonyl)amino]hexyl}carbamate;tert-butyl[(2S)-1-{[bis(4-methoxybenzyl)carbamoyl]oxy}hexan-2-yl]carbamate; methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-2-(4-methoxybenzyl)-1-(4-methoxyphenyl)-3,8-dioxo-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-({[(1S)-1-(1,3-benzodioxol-5-yl)-3-hydroxypropyl]carbamoyl}amino)hexyl bis(4-methoxybenzyl)carbamate;(2S)-2-[(tertbutoxycarbonyl)amino]hexyl dibenzylcarbamate; methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-2-benzyl-6-butyl-3,8-dioxo-1-phenyl-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(4-methylbenzyl)carbamoyl]oxy}hexan-2-yl]carbamate; methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-2-(4-methylbenzyl)-1-(4-methylphenyl)-3,8-dioxo-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(4-chlorobenzyl)carbamoyl]oxy}hexan-2-yl]carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-2-(4-chlorobenzyl)-1-(4-chlorophenyl)-3,8-dioxo-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-[(tertbutoxycarbonyl)amino]hexyl(4-bromobenzyl)(2-thienylmethyl)carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-2-(4-bromobenzyl)-6-butyl-3,8-dioxo-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-2-(4-azidoobenzyl)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-[(tert-butoxycarbonyl)amino]hexylphenyl(2-thienylmethyl)carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-2-phenyl-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(3-thienylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(3-thienyl)-2-(3-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;benzyl[(5S)-5-[(tertbutoxycarbonyl)amino]-6-{[butyl(2-thienylmethyl)carbamoyl]oxy}hexyl]carbamate;(2S)-2-[(tert-butoxycarbonyl)amino]hexylbutyl(2-thienylmethyl)carbamate;methyl(3S,7S)-3-(1,3-benzodioxol-5-yl)-7-butyl-5,11-dioxo-11-(2-thienylmethyl)-9-oxa-4,6,11-triazapentadecan-1-oate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-{[(2-methoxyethyl)(2-thienylmethyl)carbamoyl]oxy}hexyl]carbamate;(2S)-2-[(tert-butoxycarbonyl)amino]hexyl(2-methoxyethyl)(2-thienylmethyl)carbamate; methyl(9S,13S)-13-(1,3-benzodioxol-5-yl)-9-butyl-6,11-dioxo-5-(2-thienylmethyl)-2,7-dioxa-5,10,12-triazapentadecan-15-oate;(2S)-2-[({3-[(methylsulfonyl)amino]benzyl}carbamoyl)amino]hexyl(2-methoxyethyl)(2-thienylmethyl)carbamate;(2S)-2-{[(4-bromobenzyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate;(2S)-2-{[(4-azidobenzyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate;tert-butyl[(2S)-1-{[bis(2-thienylmethyl)carbamoyl]thio}hexan-2-yl]carbamate;and methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-thia-2,7,9-triazadodecan-12-oate;the four class of the compounds are selected from the group consistingofN-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}-N-(2-thienylmethyl)thiophene-2-sulfonamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}-N-(2-thienylmethyl)thiophene-2-carboxamide;2-{butyl[(2-thienylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)ethanesulfonamide;2-{[bis(2-thienylmethyl)carbamoyl](butyl)amino}-N,N-bis(2-thienylmethyl)ethanesulfonamide;N-{3-[bis(2-thienylmethyl)sulfamoyl]propyl}-N-(2-thienylmethyl)thiophene-2-sulfonamide;2-[(methylsulfonyl)(2-thienylmethyl)amino]-N,N-bis(2-thienylmethyl)ethanesulfonamide;2-{[bis(2-thienylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)ethanesulfonamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}thiophene-2-sulfonamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}-2-(2-thienyl)acetamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}thiophene-2-carboxamide;N,N-bis(2-thienylmethyl)-2-{[(2-thienylmethyl)carbamoyl]amino}ethanesulfonamide;2-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)acetamide;3-[{2-[bis(2-thienylmethyl)amino]-2-oxoethyl}(butyl)amino]-N,N-bis(2-thienylmethyl)propanamide;2-[{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}(methyl)amino]-N,N-bis(2-thienylmethyl)acetamide;2-[{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}(butyl)amino]-N,N-bis(2-thienylmethyl)acetamide;3-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)propanamide;3-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(4-methoxybenzyl)propanamide;3-({2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)propanamide;3-[{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}(methyl)amino]-N,N-bis(2-thienylmethyl)propanamide; 3-[{2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}(methyl)amino]-N,N-bis(2-thienylmethyl) propanamide;(2S)-2-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)hexanamide;(2S)-2-({2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)hexanamide;2-(acetyl{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)acetamide; and 2-(acetyl{2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienyl-methyl) acetamide;pharmaceutically acceptable salts thereof; and mixtures thereof.
 8. Themethod of claim 7, wherein, in the administering step, the antigen is ananti-cancer antigen.
 9. The method of claim 8, wherein, in theadministering step, the integrin is selected from the group consistingof α4β1, α4β7, α5β1, and αLβ2.
 10. The method of claim 8, wherein, inthe administering step, the integrin is selected from the groupconsisting of α4β1 and αLβ2.
 11. A method comprising: administering to asubject in need thereof: (A) one or more antigens, and (B) anadjuvanting amount of one or more integrin activating compounds thatincrease an immune response to the one or more antigens.
 12. The methodof claim 11, wherein the administering step include: administering theone or more antigens with the one or more integrin activating compounds.13. The method of claim 11, wherein, in the administering step, theintegrin is selected from the group consisting of α4β1, α4β7, α5β1, andαLβ2.
 14. The method of claim 11, wherein, in the administering step,the integrin is selected from the group consisting of α4β1 and αLβ2. 15.The method of claim 11, wherein, in the administering step, the one ormore integrin activating compounds comprise one or more compounds of thegeneral Formula (I):R¹-M¹-N(R²)-M²-M³-M⁴-M⁵-M⁶-R³  (I) wherein a first class of thecompounds of Formula (I) is defined by: R¹ is selected from the groupconsisting of aryl and aralkyl, R² is alkyl, aryl, or aralkyl, M¹ isCH₂, M² is CO, M³ is O, S, or NR⁶, R⁶ when present is hydrogen or loweralkyl, M⁴ is absent or CH₂, M⁵ is (CR¹¹R¹²), R¹¹ is hydrogen, R¹² isselected from the group consisting of hydrogen, NR²¹CONR²²R²³,NR²¹COR²⁴, NR²¹SO₂R²⁴, NR²¹COOR²⁴, OCOR²⁴, OR²⁴, O(CH₂CH₂O)_(s)R²⁴,COOR²⁴, alkyl, and hydroxyalkyl, s is an integer of 1 to 6, R²¹ and R²²when present are independently selected from the group consisting ofhydrogen or lower alkyl, R²³ when present is selected from the groupconsisting of hydroxyalkyl, alkoxyalkyl, alkyl, aryl, aralkyl andalkoxycarbonylalkyl, provided that when M³ is NR⁶ and M⁴ is absent, thenR²³ is not 1-(1,3-benzodioxol-5-yl)-3-ethoxy-3-oxopropyl, R²⁴ whenpresent is selected from the group consisting of alkyl, aryl, aralkyl,heterocyclyl, cycloalkyl, cycloalkylalkyl, and heterocyclylalkyl, andmixtures thereof, M⁶ is (CH₂)_(q), where q is an integer from 0 to 6, R³is selected from the group consisting of hydrogen, CONR¹³R¹⁴,NR¹⁵COOR¹⁶, NR¹⁵COR¹⁶, NR¹⁵CONR¹³R¹⁴, NR¹⁵SO₂R¹⁶, OCOR¹⁶, COOR¹⁶, OR¹⁶,SR¹⁶, heterocyclyl, hydroxyl, hydroxyalkyl, guanadino, alkyl and aryl,R¹³ and R¹⁵ when present are independently hydrogen or lower alkyl, R¹⁴and R¹⁶ when present are independently selected from the groupconsisting of hydrogen, alkyl, aryl, aralkyl, cycloalkyl, heterocyclyl,cycloalkylalkyl, and heterocyclylalkyl, R¹, R², R³, R¹², R¹⁴, R¹⁶, R²³and R²⁴ when present may independently be either unsubstituted orsubstituted with one or more substituents selected from the groupconsisting of alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl,heterocyclyl, heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy,azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl, halo,haloalkyl, haloaryl, amino, alkylamino, dialkylamino, arylamino,diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl),—NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl), —NHSO₂(aralkyl),alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino),—OCO(dialkylamino), and mixtures thereof; or wherein a second class ofthe compounds of Formula (I) is defined by: R¹ is aryl or aralkyl, R² isalkyl or aralkyl, M¹ is CH₂, M² is CO, M³ is absent or is O or CH₂, M⁴is absent or is CH₂, M⁵ is absent or is O or (CR¹¹R¹²), R¹¹ is hydrogen,R¹² is selected from the group consisting of hydrogen, NR²¹CONR²²R²³,NR²¹COR²⁴, NR²¹SO₂R²⁴ and NR²¹COOR²⁴, R²¹ and R²² each of which, whenpresent is independently selected from the group of hydrogen and loweralkyl, R²³ and R²⁴, each of which, when present is independentlyselected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, M⁶ isselected from the group consisting of (CH₂)_(q),(CH₂)_(q)—CH═CH—(CH₂)_(r), (CH₂)_(q)-arylene-(CH₂)_(r) and(CH₂CH₂O)_(q), where q and r are independently integers from 0 to 6, R³is CONR¹³R¹⁴, R¹³ and R¹⁴, each of which, when present is independentlyselected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, and R¹,R², R¹³, R¹⁴, R²³ and R²⁴, when present, independently either areunsubstituted or are substituted with one or more substituents selectedfrom the group consisting of alkyl, aryl, aralkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl,hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl,alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino,arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl),—NHCO(haloalkyl), —NHSO₂(alkyl), —NHSO₂(aryl), —NHSO₂(aralkyl),alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino),—OCO(dialkylamino), and mixtures thereof; or wherein a third class ofthe compounds of Formula (I) is defined by: R¹ is aryl or aralkyl, R² isalkyl or aralkyl, M¹ is CH₂, M² is SO₂ or CO, M³ is absent or is CH₂, M⁴is absent or is CH₂, M⁵ is absent or is (CR¹¹R¹²), R¹¹, when present, ishydrogen, R¹², when present, is selected from the group consisting ofhydrogen, alkyl, NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴ and NR²¹COOR²⁴,R²¹ and R²², each of which when present, is independently selected fromthe group of hydrogen, lower alkyl, and aralkyl, R²³ and R²⁴, each ofwhich, when present is independently selected from the group consistingof hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl,heterocyclylalkyl and aralkyl, M⁶ is (CH₂)_(q), or NR³⁴(CH₂)_(q), q isan integer from 0 to 6, R³⁴, when present, is selected form the groupconsisting of alkyl, aralkyl, COR³⁵, and SO₂R³⁵, R³⁵ when present, isselected form the group consisting of alkyl, aryl, and aralkyl, and R³is selected from the group consisting of CONR¹³R¹⁴, SO₂NR¹³R¹⁴,NR¹⁵COOR¹⁶, NR¹⁵COR¹⁶, NR¹⁵CONR¹³R¹⁴, and NR¹⁵SO₂R¹⁶, R¹³ and R¹⁴, eachof which, when present, is independently selected from the groupconsisting of hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl,heterocyclyl, heterocyclylalkyl and aralkyl, R¹⁵ and R¹⁶, each of whichwhen present, is independently selected from the group of hydrogen,lower alkyl, and aralkyl, R¹, R², R¹³, R¹⁴, R¹⁵, R¹⁶, R²³, R²⁴, R³⁴ andR³⁵, when present, either are unsubstituted or are substituted with oneor more substituents selected from the group consisting of alkyl, aryl,aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl,heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl,aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino,alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl),—NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO₂(alkyl),—NHSO₂(aryl), —NHSO₂(aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl,—OCO(alkylamino), and —OCO(dialkylamino), with the proviso that when M²is CO, then M⁶ is NR³⁴(CH₂)_(q), q is not 0; or wherein a four class ofthe compounds of Formula (I) is defined by: R¹ is alkyl, aryl oraralkyl, R² is selected from the group consisting of aralkyl and alkyl,provided that when R¹ is alkyl, R² is aralkyl, M¹ is CO or SO₂, providedthat when M¹ is SO₂ and R¹ is phenyl, 4-methylphenyl or2,4,6-trimethylphenyl, R² is not alkyl, 2-phenethyl, benzyl, or2-methoxy-2-oxoethyl, and when M′ is CO and R′ is 2-furyl, 4-pyridyl, or3,5-dinitrophenyl, R² is not alkyl, benzyl or 2-(1H-indol-2-yl)ethyl, M²is absent or CH₂, M³ and M⁴ are absent, M⁵ is (CR¹¹R¹²), R¹¹ ishydrogen, R¹² is selected from the group consisting of hydrogen,NR²¹CONR²²R²³, NR²¹COR²⁴, NR²¹SO₂R²⁴, NR²¹COOR²⁴, CONR²²R²³, COOR²⁴,O(CH₂CH₂O)_(s)R²⁴, hydroxyalkyl, and alkoxyalkyl, R²¹, and R²², whenpresent, are independently selected from the group consisting ofhydrogen and C₁-C₆ alkyl, and R²³ and R²⁴, each of which, when present,is independently selected from the group consisting of hydrogen, alkyl,aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl andaralkyl, and s is an integer of 1 to 6, M⁶ is (CH₂)_(q), q is an integerof 0 to 6, R³ is selected from the group consisting of NR¹⁵COOR¹⁶,NR¹⁵COR¹⁶, NR¹⁵CONR¹³R¹⁴, and NR¹⁵SO₂R¹⁶, and R¹³ when present, isindependently selected from the group consisting of hydrogen and C₁-C₆alkyl, and R¹⁴, R¹⁵, and R¹⁶ each of which, when present, areindependently selected from the group consisting of hydrogen, alkyl,aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl andaralkyl, and R¹, R², R³, R¹², R¹⁴, R¹⁵, R¹⁶, R²³, and R²⁴, when present,independently either are unsubstituted or are substituted with one ormore substituents selected from the group consisting of alkyl, aryl,aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl,heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl,aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino,alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl),—NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO₂(alkyl),—NHSO₂(aryl), —NHSO₂(aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl,—OCO(alkylamino) and —OCO(dialkylamino), or pharmaceutically acceptablesalts thereof, or mixtures thereof.
 16. The method of claim 15, wherein,in the administering step, R¹M¹, R², R¹³, and R¹⁴ are independentlyselected from the group consisting of 2-thienylmethyl, 3-methoxybenzyl,4-methoxybenzyl, 2-(2-thienyl)ethyl, pyridin-4-ylmethyl, andpyridin-3-ylmethyl.
 17. The method of claim 16, wherein, in theadministering step: the first class of the compounds are selected fromthe group consisting ofN,N,N′,N′-tetrakis(2-thienylmethyl)pentanediamide;N-(3-methoxybenzyl)-N,N′,N′-tris(2-thienylmethyl)pentanediamide;N,N,N′-tris(2-thienylmethyl)pentanediamide;N′-[2-(2-thienyl)ethyl]-N,N-bis(2-thienylmethyl)pentanediamide;N-[2-(2-thienyl)ethyl]-N,N′,N′-tris(2-thienylmethyl)pentanediamide;N,N-bis(pyridin-4-ylmethyl)-N′,N′-bis(2-thienylmethyl)pentanediamide;N,N-bis(pyridin-3-ylmethyl)-N′,N′-bis(2-thienylmethyl)pentanediamide;N,N-bis(3-methoxybenzyl)-N′,N′-bis(2-thienylmethyl)pentanediamide;N,N,N′,N′-tetrakis(4-methoxybenzyl)pentanediamide;N,N,N′,N′-tetrakis(2-thienylmethyl)hexanediamide;N,N,N′,N′-tetrakis(4-methoxybenzyl) hexanediamide;N,N,N′,N′-tetrakis(3-methoxybenzyl)hexanediamide; N,N,N′,N′-tetrakis(2-thienylmethyl) heptanediamide;2,2′-(1,3-phenylene)bis[N,N-bis(2-thienylmethyl)acetamide];N,N,N′,N′-tetrakis(4-methoxybenzyl)heptanediamide; N,N,N′,N′-tetrakis(2-thienylmethyl)octanediamide; (3E)-N,N,N′,N′-tetrakis(2-thienylmethyl)hex-3-enediamide;2,2′-oxybis[N,N-bis(2-thienylmethyl)acetamide];3-oxo-1-(2-thienyl)-2-(2-thienylmethyl)-4,7,10-trioxa-2-azadodecan-12-ylbis(2-thienylmethyl)carbamate; N,N,N′,N′-tetrakis(4-methoxybenzyl)succinamideethane-1,2-diyl bis[bis(2-thienylmethyl)carbamate];N,N,N′,N′-tetrakis (4-methoxybenzyl)octanediamide;N,N,N′,N′-tetrakis(2-thienylmethyl) pyridine-3,5-dicarboxamide;N,N,N′,N′-tetrakis (2-thienylmethyl)pyridine-2,6-dicarboxamide;N,N,N′,N′-tetrakis (2-thienylmethyl)pyridine-2,4-dicarboxamide;2,2′-(1,4-phenylene) bis[N,N-bis(2-thienylmethyl)acetamide]; 8-{2-[bis(2-thienylmethyl)amino]-2-oxoethoxy}-N,N-bis(2-thienylmethyl)quinoline-2-carboxamide;N,N′-bis(4-methoxybenzyl)-N,N′-bis(2-thienylmethyl)hexanediamide; andtert-butyl{(2S)-1,6-bis[bis(2-thienylmethyl)amino]-1,6-dioxohexan-2-yl}carbamate;the second class of the compounds are selected from the group consistingof 1,2-bis(bis(thiophen-2-ylmethyl)carbamate)ethane;1,2-bis(bis(3-methyloxybenzyl)carbamate)ethane;1,2-bis(bis(4-methyloxybenzyl)carbamate)ethane;1,2-bis((thiophen-2-ylmethyl)(3-methyloxybenzyl)carbamate)ethane;1,2-bis((thiophen-2-ylmethyl)(4-methyloxybenzyl)carbamate)ethane;1,2-bis((3-methyloxybenzyl)(4-methyloxybenzyl)carbamate)ethane;1,5-bis(bis(thiophen-2-ylmethyl)carbamate)-3-oxapentane;1,5-bis(bis(3-methyloxybenzyl)carbamate)-3-oxapentane;1,5-bis(bis(4-methyloxybenzyl)carbamate)-3-oxapentane;1,5-bis((thiophen-2-ylmethyl)(3-methyloxybenzyl)carbamate)-3-oxapentane;1,5-bis((thiophen-2-ylmethyl)(4-methyloxybenzyl)carbamate)-3-oxapentane;1,5-bis((3-methyloxybenzyl)(4-methyloxybenzyl)carbamate)-3-oxapentane;(ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl)bis(bis(thiophen-2-ylmethyl)carbamate)or 1,8-bis(bis(thiophen-2-ylmethyl)carbamate)-3,6-dioxaoctane;1,8-bis(bis(3-methyloxybenzyl)carbamate)-3,6-dioxaoctane;1,8-bis(bis(4-methyloxybenzyl)carbamate)-3,6-dioxaoctane;1,8-bis((thiophen-2-ylmethyl)(3-methyloxybenzyl)carbamate)-3,6-dioxaoctane;1,8-bis((thiophen-2-ylmethyl)(4-methyloxybenzyl)carbamate)-3,6-dioxaoctane;1,8-bis((3-methyloxybenzyl)(4-methyloxybenzyl)carbamate)-3,6-dioxaoctane;1,11-bis(bis(thiophen-2-ylmethyl)carbamate)-3,6,9-trioxaundecane;1,11-bis(bis(3-methyloxybenzyl)carbamate)-3,6,9-trioxaundecane;1,11-bis(bis(4-methyloxybenzyl)carbamate)-3,6,9-trioxaundecane;1,11-bis((thiophen-2-ylmethyl)(3-methyloxybenzyl)carbamate)-3,6,9-trioxaundecane;1,11-bis((thiophen-2-ylmethyl)(4-methyloxybenzyl)carbamate)-3,6,9-trioxaundecane;1,11-bis((3-methyloxybenzyl)(4-methyloxybenzyl)carbamate)-3,6,9-trioxaundecane;1,14-bis(bis(thiophen-2-ylmethyl)carbamate)-3,6,9,12-tetraoxatetradecane;1,14-bis(bis(3-methyloxybenzyl)carbamate)-3,6,9,12-tetraoxatetradecane;1,14-bis(bis(4-methyloxybenzyl)carbamate)-3,6,9,12-tetraoxatetradecane;1,14-bis((thiophen-2-ylmethyl)(3-methyloxybenzyl)carbamate)-3,6,9,12-tetraoxatetradecane;1,14-bis((thiophen-2-ylmethyl)(4-methyloxybenzyl)carbamate)-3,6,9,12-tetraoxatetradecane;1,14-bis((3-methyloxybenzyl)(4-methyloxybenzyl)carbamate)-3,6,9,12-tetraoxatetradecane;1,17-bis(bis(thiophen-2-ylmethyl)carbamate)-3,6,9,12,15-pentaoxatetradecane;1,17-bis(bis(3-methyloxybenzyl)carbamate)-3,6,9,12,15-pentaoxatetradecane;1,17-bis(bis(4-methyloxybenzyl)carbamate)-3,6,9,12,15-pentaoxatetradecane;1,17-bis((thiophen-2-ylmethyl)(3-methyloxybenzyl)carbamate)-3,6,9,12,15-pentaoxatetradecane;1,17-bis((thiophen-2-ylmethyl)(4-methyloxybenzyl)carbamate)-3,6,9,12,15-pentaoxatetradecane;and1,17-bis((3-methyloxybenzyl)(4-methyloxybenzyl)carbamate)-3,6,9,12,15-pentaoxatetradecane;the third class of the compounds are selected from the group consistingof methyl(6S,10S)-1-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10R)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-7-methyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-9-methyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;ethyl(6S,10R)-1-(1,3-benzodioxol-5-yl)-6-butyl-7-methyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(10S)-10-(1,3-benzodioxol-5-yl)-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-2-methyl-3,8-dioxo-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-{[(1,3-benzodioxol-5-ylmethyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate; methyl(6S,10S)-6-butyl-3,8-dioxo-10-phenyl-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-({[(1S)-1-(1,3-benzodioxol-5-yl)-3-hydroxypropyl]carbamoyl}amino)hexylbis(2-thienylmethyl)carbamate;(2S)-2-[(benzylcarbamoyl)amino]hexylbis(2-thienylmethyl)carbamate;(2S)-2-[(morpholin-4-ylcarbonyl)amino]hexylbis(2-thienylmethyl)carbamate;(2S)-2-{[(3-methoxypropyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate;(2S)-2-{[(2-methoxyethyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate;tert-butyl[(2S)-1-{[bis(2-thienylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;(2S)-2-[(tert-butylcarbamoyl)amino]hexyl bis(2-thienylmethyl)carbamate;(2S)-2-[(isopropylcarbamoyl)amino]hexylbis(2-thienylmethyl)carbamate;(2S)-2-[(methylcarbamoyl)amino]hexyl bis(2-thienylmethyl)carbamate;tert-butyl[(2R)-1-{[bis(2-thienylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;benzyl{(5S)-6-{[bis(2-thienylmethyl)carbamoyl]oxy}-5-[(tert-butoxycarbonyl)amino]hexyl}carbamate;methyl(9S,13S)-13-(1,3-benzodioxol-5-yl)-9-({[bis(2-thienylmethyl)carbamoyl]oxy}methyl)-3,11-dioxo-1-phenyl-2-oxa-4,10,12-triazapentadecan-15-oate;(2S)-2-acetamidohexylbis(2-thienylmethyl)carbamate;methyl(6R,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6R,10R)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl{[bis(2-thienylmethyl)carbamoyl](methyl)amino}acetate;methyl{[bis(2-thienylmethyl)carbamoyl](butyl)amino}acetate;tert-butyl{[bis(2-thienylmethyl)carbamoyl](butyl)amino}acetate;benzyl{(5S)-6-{[bis(4-methoxybenzyl)carbamoyl]oxy}-5-[(tert-butoxycarbonyl)amino]hexyl}carbamate;tert-butyl[(2S)-1-{[bis(4-methoxybenzyl)carbamoyl]oxy}hexan-2-yl]carbamate; methyl(6S,10S)-0-(1,3-benzodioxol-5-yl)-6-butyl-2-(4-methoxybenzyl)-1-(4-methoxyphenyl)-3,8-dioxo-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-({[(1S)-1-(1,3-benzodioxol-5-yl)-3-hydroxypropyl]carbamoyl}amino)hexyl bis(4-methoxybenzyl)carbamate;(2S)-2-[(tertbutoxycarbonyl)amino]hexyl dibenzylcarbamate; methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-2-benzyl-6-butyl-3,8-dioxo-1-phenyl-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(4-methylbenzyl)carbamoyl]oxy}hexan-2-yl]carbamate; methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-2-(4-methylbenzyl)-1-(4-methylphenyl)-3,8-dioxo-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(4-chlorobenzyl)carbamoyl]oxy}hexan-2-yl]carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-2-(4-chlorobenzyl)-1-(4-chlorophenyl)-3,8-dioxo-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-[(tertbutoxycarbonyl)amino]hexyl(4-bromobenzyl)(2-thienylmethyl)carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-2-(4-bromobenzyl)-6-butyl-3,8-dioxo-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;methyl(6S,10S)-2-(4-azidoobenzyl)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;(2S)-2-[(tert-butoxycarbonyl)amino]hexylphenyl(2-thienylmethyl)carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-2-phenyl-1-(2-thienyl)-4-oxa-2,7,9-triazadodecan-12-oate;tert-butyl[(2S)-1-{[bis(3-thienylmethyl)carbamoyl]oxy}hexan-2-yl]carbamate;methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(3-thienyl)-2-(3-thienylmethyl)-4-oxa-2,7,9-triazadodecan-12-oate;benzyl[(5S)-5-[(tertbutoxycarbonyl)amino]-6-{[butyl(2-thienylmethyl)carbamoyl]oxy}hexyl]carbamate;(2S)-2-[(tert-butoxycarbonyl)amino]hexylbutyl(2-thienylmethyl)carbamate;methyl(3S,7S)-3-(1,3-benzodioxol-5-yl)-7-butyl-5,11-dioxo-11-(2-thienylmethyl)-9-oxa-4,6,11-triazapentadecan-1-oate;benzyl[(5S)-5-[(tert-butoxycarbonyl)amino]-6-{[(2-methoxyethyl)(2-thienylmethyl)carbamoyl]oxy}hexyl]carbamate;(2S)-2-[(tert-butoxycarbonyl)amino]hexyl(2-methoxyethyl)(2-thienylmethyl)carbamate; methyl(9S,13S)-13-(1,3-benzodioxol-5-yl)-9-butyl-6,11-dioxo-5-(2-thienylmethyl)-2,7-dioxa-5,10,12-triazapentadecan-15-oate;(2S)-2-[({3-[(methylsulfonyl)amino]benzyl}carbamoyl)amino]hexyl(2-methoxyethyl)(2-thienylmethyl)carbamate;(2S)-2-{[(4-bromobenzyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate;(2S)-2-{[(4-azidobenzyl)carbamoyl]amino}hexylbis(2-thienylmethyl)carbamate;tert-butyl[(2S)-1-{[bis(2-thienylmethyl)carbamoyl]thio}hexan-2-yl]carbamate;and methyl(6S,10S)-10-(1,3-benzodioxol-5-yl)-6-butyl-3,8-dioxo-1-(2-thienyl)-2-(2-thienylmethyl)-4-thia-2,7,9-triazadodecan-12-oate;the four class of the compounds are selected from the group consistingofN-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}-N-(2-thienylmethyl)thiophene-2-sulfonamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}-N-(2-thienylmethyl)thiophene-2-carboxamide;2-{butyl[(2-thienylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)ethanesulfonamide;2-{[bis(2-thienylmethyl)carbamoyl](butyl)amino}-N,N-bis(2-thienylmethyl)ethanesulfonamide;N-{3-[bis(2-thienylmethyl)sulfamoyl]propyl}-N-(2-thienylmethyl)thiophene-2-sulfonamide;2-[(methylsulfonyl)(2-thienylmethyl)amino]-N,N-bis(2-thienylmethyl)ethanesulfonamide;2-{[bis(2-thienylmethyl)carbamoyl]amino}-N,N-bis(2-thienylmethyl)ethanesulfonamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}thiophene-2-sulfonamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}-2-(2-thienyl)acetamide;N-{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}thiophene-2-carboxamide;N,N-bis(2-thienylmethyl)-2-{[(2-thienylmethyl)carbamoyl]amino}ethanesulfonamide;2-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)acetamide;3-[{2-[bis(2-thienylmethyl)amino]-2-oxoethyl}(butyl)amino]-N,N-bis(2-thienylmethyl)propanamide;2-[{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}(methyl)amino]-N,N-bis(2-thienylmethyl)acetamide;2-[{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}(butyl)amino]-N,N-bis(2-thienylmethyl)acetamide;3-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)propanamide;3-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(4-methoxybenzyl)propanamide;3-({2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)propanamide;3-[{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}(methyl)amino]-N,N-bis(2-thienylmethyl)propanamide; 3-[{2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}(methyl)amino]-N,N-bis(2-thienylmethyl) propanamide;(2S)-2-({2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)hexanamide;(2S)-2-({2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)hexanamide;2-(acetyl{2-[bis(2-thienylmethyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienylmethyl)acetamide; and 2-(acetyl{2-[bis(4-methoxybenzyl)sulfamoyl]ethyl}amino)-N,N-bis(2-thienyl-methyl) acetamide;pharmaceutically acceptable salts thereof; and mixtures thereof.
 18. Themethod of claim 17, wherein, in the administering step, the one or moreantigens are anti-cancer antigens.
 19. The method of claim 18, wherein,in the administering step, the integrin is selected from the groupconsisting of α4β1, α4β7, α5β1, and αLβ2.
 20. The method of claim 18,wherein, in the administering step, the integrin is selected from thegroup consisting of α4β1 and αLβ2.